Sample records for rennet-induced casein gels

During ageing of casein or skim milk gels, structural changes take place that affect gel parameters, such as pore size and storage modulus. These changes can be explained in terms of rearrangements of the gel network at various length scales. In this paper, rheological experiments on rennet-induced

The rheological properties of rennet-induced skim milk gels, which are viscoelastic materials, were studied under various conditions.Dynamic and stress relaxation experiments were performed at small deformations of the gel network, whereas constant stress (creep) experiments were performed at large

The effects of high-pressure homogenisation (HPH) in the pressure range 100-300 MPa on the gel formation and rheological properties of rennet-induced skim milk (0.08%, fat, w/w) and standardised milk (3.60% fat, w/w) gels at pH 6.60 were studied. The average casein micelle size in skim milk was significantly reduced and the gel formation time decreased when skim milk was subjected to the pressures of 200 and 300 MPa. The storage modulus of rennet-induced skim milk gels at 2700 s after rennet addition was higher for samples homogenised at higher pressures, which contained smaller casein particles. HPH had little effect on the large deformation properties of rennet-induced skim milk gels. The gel formation time of renneted standardised milk was significantly reduced as a result of HPH, while the storage modulus of rennet-induced milk gels 2700 s after rennet addition increased with increasing homogenising pressure. The apparent fracture stress was slightly higher for standardised milk gels formed from HPH-treated milk, whereas the apparent strain at fracture was lower, than that of unhomogenised milk. In conclusion, HPH treatment influenced gel formation processes of skim milk and its small-deformation rheological properties, mainly through modification of casein micelles. HPH also significantly affected the gel formation process of standardised milk gels and its rheological properties as a result of an increase in volume fraction of aggregating particles, while the particle size was of lesser importance.

Optimizing cheese yield and quality is of central importance to cheese manufacturing. The yield is associated with the time it takes before the gel has an optimal consistency for further processing, and it is well known that gel formation differs between individual milk samples. By identifying...... genomic regions affecting traits related to rennet-induced gelation, the aim of this study was to identify potential candidate genes affecting these traits. Hence, rennet-induced gelation, including rennet coagulation time, gel strength, and yield stress, was measured in skim milk samples collected from...

The syneresis behaviour of rennet-induced skimmilk gels was studied at closely controlled conditions. The origin of the endogenous syneresis pressure and changes therein and in the permeability of the gel during syneresis are discussed. The latter appears to be of predominant importance for the oned

Rennet-inducedcaseingels made from skim milk were studied rheologically. A scaling model or framework for describing the rheological behavior of gels is discussed and used for classification of the structure of caseingels. There are two main parameters in the model that describe the number of def

Confocal laser scanning microscopy (CLSM) was successfully used to observe the effect of milk processing on the size and the morphology of the milk fat globule in raw milk, raw ultrafiltered milk, and standardized and pasteurized milk prepared for cheese manufacture (cheese-milk) and commercial pasteurized and homogenized milk. Fat globule size distributions for the milk preparations were analyzed using both image analysis and light scattering and both measurements produced similar data trends. Changes to the native milk fat globule membrane (MFGM) were tracked using a MFGM specific fluorescent stain that allowed MFGM proteins and adsorbed proteins to be differentiated on the fat globule surface. Sodium dodecyl sulfate polyacrylamide gel electrophoresis confirmed the identity of native MFGM proteins isolated from the surface of fat globules within raw, UF retentate, and cheese-milk preparations, whereas only casein was detected on the surface of fat globules in homogenized milk. The microstructure, porosity, and gel strength of the rennetinducedgel made from raw milk and cheese-milk was also found to be comparable and significantly different to that made from homogenized milk. Our results highlight the potential use of CLSM as a tool to observe the structural details of the fat globule and associated membrane close to its native environment.

lactose, to obtain varying levels of micellar calcium and phosphorus but constant value of pH, serum and free calcium, and serum phosphorus. Bovine chymosin was added to the skim milk samples after dialysis and microstructural and rheological properties during gel formation were recorded at 30°C. Samples...... of rennet gel formation. The protein network of rennet gels after dialysis was more compact with many aggregates as demineralization decreased. The small protein particles are able to increase the potential connection points among proteins, support particle fusion and cause a compact structure....... after dialysis needed approximately 30min after the addition of chymosin to form rennet gels. In addition, low micellar calcium and phosphorus values were both found to correlate with slightly less time for the gels to be formed. This information highlights the importance of CCP in the primary phase...

Concentrating milk is a common unit operation in the dairy industry. With the reduction of water, the particles interact more frequently with each other and the functionality of the casein micelles may depend on the interactions occurring during concentration. The objective of this research was to investigate the effect of concentration on the renneting properties of the casein micelles by comparing 2 concentration methods: ultrafiltration and osmotic stressing. Both methods selectively concentrate the protein fraction of milk, while the composition of the soluble phase is unaltered. To evaluate possible differences in the rearrangements of the casein micelles during concentration, renneting properties were evaluated with or without the addition of soluble caseins, added either before or after concentration. The results indicate that casein micelles undergo rearrangements during concentration and that shear during membrane filtration may play a role in affecting the final properties of the milk.

The effects of gelation temperature (20, 30 or 40 °C), assay temperature, concentration of glucono-δ-lactone (GDL) added, and NaCl concentration on the rheological properties of acid caseingels were studied at small and large deformations. Gels prepared at a high incubation temperature had very low

The influence of the ingredients of acidified Na caseinate-xanthan-sucrose gels on thermophysical properties and syneresis of the gels was studied. Sucrose concentration affected all of the gel equilibrium properties and the rate of syneresis. The positive effect of sucrose on syneresis and unfrozen water (UFW) values was attributed to different effects. The amount of UFW was governed mainly by the colligative properties of sucrose whereas the equilibrium syneresis behaviour was associated with the changes in network dynamics caused by the kosmotropic properties of sucrose. The latter could enhance xanthan-sucrose association or favour xanthan-protein interactions.

The effects of addition of alpha-casein (alpha-CN) to dried egg white (DEW) were investigated by measuring transparency, hardness, and water-holding capacity (WHC) of the heat-induced gels. A DEW concentration of 8% (w/w) was required for formation of a self-supporting gel following heating at 80 degrees C for 20 min at pH 7. Solutions of alpha-CN, even up to a protein concentration of 12% (w/w), did not gel under the same conditions. The addition of alpha-CN (0.5-4%) to 8% DEW caused the increase in gel hardness gels, as compared with DEW gels alone at a total amount of protein concentrations, and the mixed gels became transparent with the increase of added alpha-CN concentrations. The 10% mixed protein solutions of alpha-CN (3-6%) and DEW (4-7%) formed transparent gels, although each protein did not gel individually at their protein concentrations. Mixture with 2:8 mixing ratio of alpha-CN to DEW at a total protein concentration of 10% showed synergistic effects in improving DEW gel properties above pH 7 and below 25 mM NaCl. The improvements (hardness, transparency, and WHC) of DEW gel by alpha-CN seem to be caused mainly by the inhibition of alpha-CN against heat coagulation of DEW protein.

Full Text Available The aim of this work was to study the effect of xanthan gum (XG on the gelation process of bovine sodium caseinate (NaCAS induced by acidification with glucono-δ-lactone (GDL and on the mixed acid gel microstructure. Before GDL addition, segregative phase separation was observed in all the NaCAS-XG mixtures evaluated. The gelation process was analyzed by using a fractional factorial experimental design. The images of the microstructure of the mixed acid gels were obtained by conventional optical microscopy and the mean diameter of the interstices was determined. Both the elastic character and the microstructure of the gels depended on the concentrations of XG added. As XG concentration increased, the kinetics of the gelation process was modified and the degree of compactness and elasticity component of the gel network increased. The microstructure of gels depends on the balance among thermodynamic incompatibility, protein gelation and NaCAS-XG interactions.

The effects of sodium caprate on the gelation of β-lactoglobulin B and a β-lactoglobulin B/β-casein mixture at ambient temperature were investigated using ultrasonic spectroscopy and rheology. A 12% β-lactoglobulin B solution gelled in the presence of 3.6% sodium caprate. Conversely, sodium caprate did not induce the formation of a gel when β-casein was in isolation, regardless of the protein concentration. Although a 6% β-lactoglobulin B/1.8% sodium caprate solution did not form a gel, a gel was formed when 6% β-casein was added to a mixture containing 6% β-lactoglobulin B and 3.6% sodium caprate. This gel showed comparable rheological properties to that of a gel containing 12% β-lactoglobulin B. The results clearly indicated that β-casein aids in the gelation of a β-lactoglobulin B/sodium caprate mixture, when the concentration of β-lactoglobulin B is insufficient to allow for gelation. It appears that β-casein self-aggregation is also inhibited. Therefore, it could be concluded that β-casein can be used as a texture modifier for β-lactoglobulin gelation induced by sodium caprate.

Syneresis of caseingels made by acidification with glucono-δ-lactone (GDL) was studied in relation to gel structure as derived from permeametry and confocal scanning laser microscopy (CSLM). Gels made at 40 °C exhibited ‘spontaneous syneresis’ with wheying-off almost immediately after gelation, whi

Rennet-induced gelation crucially impacts cheese structure. In this study, effects of the size and stability of native fat globules on the kinetics of rennet-induced coagulation were revealed by determining the caseinomacropeptide release rate and rheological properties of milk. Moreover, the mobility and stability of fat globules during renneting was revealed using diffusing wave spectroscopy and confocal laser scanning microscopy. By use of a 2-stage gravity separation combined centrifugation scheme, native fat globules were selectively separated into small (SFG, D4,3 = 1.87 ± 0.02 μm) and large fat globules (LFG, D4,3 = 5.65 ± 0.03 μm). The protein and fat content of SFG and LFG milk were then standardized to 3.2 g/100 mL and 1.2 g/100 mL, respectively. The milk containing different sized globules were then subjected to renneting experiments in the laboratory. Reduction of globule size accelerated the aggregation of casein micelles during renneting, giving a shorter gelation time and earlier 1/l(*) change. The gel produced from LFG milk was broken due to coalescent fat globules and generated coarser gel strands compared to the finer strands formed with SFG milk. Structural differences were also confirmed with a higher final storage modulus of the curd made from SFG milk than that from the LFG. In conclusion, the size of fat globules affects the aggregation of casein micelles. Moreover, fat globule coalescence and creaming during renneting, also affects the structure of the rennet gel. A better understanding of the size of globules effect on milk gelation could lead to the development of cheese with specific properties.

The economic output of the dairy industry is to a great extent dependent on the processing of milk into other milk-based products such as cheese. The yield and quality of cheese are dependent on both the composition and technological properties of milk. The objective of this study was to evaluate the importance and effects of casein (CN) micelle size and milk composition on milk gelation characteristics in order to evaluate the possibilities for enhancing gelation properties through breeding. Milk was collected on 4 sampling occasions at the farm level in winter and summer from dairy cows with high genetic merit, classified as elite dairy cows, of the Swedish Red and Swedish Holstein breeds. Comparisons were made with milk from a Swedish Red herd, a Swedish Holstein herd, and a Swedish dairy processor. Properties of CN micelles, such as their native and rennet-induced CN micelle size and their zeta-potential, were analyzed by photon correlation spectroscopy, and rennet-induced gelation characteristics, including gel strength, gelation time, and frequency sweeps, were determined. Milk parameters of the protein, lipid, and carbohydrate profiles as well as minerals were used to obtain correlations with native CN micelle size and gelation characteristics. Milk pH and protein, CN, and lactose contents were found to affect milk gelation. Smaller native CN micelles were shown to form stronger gels when poorly coagulating milk was excluded from the correlation analysis. In addition, milk pH correlated positively, whereas Mg and K correlated negatively with native CN micellar size. The milk from the elite dairy cows was shown to have good gelation characteristics. Furthermore, genetic progress in relation to CN micelle size was found for these cows as a correlated response to selection for the Swedish breeding objective if optimizing for milk gelation characteristics. The results indicate that selection for smaller native CN micelles and lower milk pH through breeding would

The objective of this study was to investigate the impact of natural variations in aS1-casein and b-casein composition of milk on chymosin-induced hydrolysis of these caseins in milk gels and in sodium caseinate solutions. At 50% casein degradation, 15% more of aS1-casein with eight phosphate groups

Full Text Available Technological operations applied during curd processing influence syneresis and total solids content of cheese. Syneresis is not a simple physical process representing whey segregation due to curd contractions. Numerous factors can influence the process of syneresis. The aim of this work was to investigate the influence of various parameters (pH, quantity of CaCl2 added, temperature of coagulation and heat treatment on induced syneresis. Reconstituted instant skim milk (control samples and reconstituted instant skim milk heated at 87ºC for 10 min (experimental samples were coagulated at 30ºC and 35ºC, and pH of 5.8 and 6.2 with 100, 200 and 400 mg/l of CaCl2 added. According to our results, these parameters had significant influence on nitrogen content of serum as well as on the distribution of nitrogen matter from gel into sera. Due to the formation of coaggregates the best rheological properties of gel were obtained for experimental samples coagulated with 400 mg/l of CaCl2 added at pH 5.8 and temperature of 35ºC.

Recent studies on gel formation and rheology of milk gels are reviewed. A distinction is made between gels formed by aggregated casein, gels of `pure` whey proteins and gels in which both casein and whey proteins contribute to their properties. For casein' whey protein mixtures, it has been shown th

Recent studies on gel formation and rheology of milk gels are reviewed. A distinction is made between gels formed by aggregated casein, gels of `pure` whey proteins and gels in which both casein and whey proteins contribute to their properties. For casein' whey protein mixtures, it has been shown

Casein micelles are porous colloidal particles, constituted of casein molecules, water, and minerals. The vulnerability of the supramolecular structure of casein micelles face to changes in the environmental conditions restrains their applications in other domains besides food. Thus, redesigning casein micelles is a challenge to create new functionalities for these biosourced particles. The objective of this work was to create stable casein microgels from casein micelles using a natural cross-linker, named genipin. Suspensions of purified casein micelles (25 g L(-1)) were mixed with genipin solutions to have final concentrations of 5, 10, and 20 mM genipin. Covalently linked casein microgels were formed via cross-linking of lysyl and arginyl residues of casein molecules. The reacted products exhibited blue color. The cross-linking reaction induced gradual changes on the colloidal properties of the particles. The casein microgels were smaller and more negatively charged and presented smoother surfaces than casein micelles. These results were explained based on the cross-linking of free NH2 present in an external layer of κ-casein. Light scattering and rheological measurements showed that the reaction between genipin and casein molecules was intramicellar, as one single population of particles was observed and the values of viscosity (and, consequently, the volume fraction of the particles) were reduced. Contrary to the casein micelles, the casein microgels were resistant to the presence of dissociating agents, e.g., citrate (calcium chelating) and urea, but swelled as a consequence of internal electrostatic repulsion and the disruption of hydrophobic interactions between protein chains. The casein microgels did not dissociate at the air-solution interface and formed solid-like interfaces rather than a viscoelastic gel. The potential use of casein microgels as adaptable nanocarriers is proposed in the article.

The water holding capacity of caseingels was investigated by measuring the swelling and de-swelling under a variety of conditions of temperature and salt concentration. Transglutaminase cross-linked sodium caseinate (15% w/w) gels will swell in good solvents or shrink in poor solvents until an equi

In recent years there has been an increasing interest in pure casein fractions, particularly β-casein due to its physiochemical properties as well as its bio- and techno-functional properties. A range of methods has been developed for the fractionation of casein into its individual proteins. The

Full Text Available Context: Demineralization of tooth by erosion is caused by frequent contact between the tooth surface and acids present in soft drinks. Aim: The present study objective was to evaluate the remineralization potential of casein-phosphopeptide-amorphous calcium phosphate (CPP-ACP paste, 1.23% acidulated phosphate fluoride (APF gel and iron supplement on dental erosion by soft drinks in human primary and permanent enamel using atomic force microscopy (AFM. Materials and Methods: Specimens were made from extracted 15 primary and 15 permanent teeth which were randomly divided into three treatment groups: CPP-ACP paste, APF gel and iron supplement. AFM was used for baseline readings followed by demineralization and remineralization cycle. Results and Statistics: Almost all group of samples showed remineralization that is a reduction in surface roughness which was higher with CPP-ACP paste. Statistical analysis was performed using by one-way ANOVA and Mann-Whitney U-test with P < 0.05. Conclusions: It can be concluded that the application of CPP-ACP paste is effective on preventing dental erosion from soft drinks.

Bovine milk contains 3-4 % protein and almost 80% of the milk protein fraction consist of four caseins; αs1-casein, β-casein, αs2-casein and κ-casein. Most of the caseins in milk are assembled in casein micelles, which consist of several thousands of indi

Aim of this study was to investigate the digestion of transglutaminase cross-linked caseinate (XLC) versus native caseinate (NC) in solution and in cheese spread under digestive conditions for adults and children mimicked in a gastrointestinal model. Samples were collected for gel electrophoresis an

The concentrated (dispersed phase 50-70 wt%) composition space of Na-caseinate, a family of milk proteins, stabilised emulsions was investigated for three different oils: soybean oil, palm olein and tetradecane with pH 6.8 phosphate buffer continuous phase. The variation of emulsion stability and microstructure were explored using static light scattering, diffusion nuclear magnetic resonance, cryo-scanning electron microscopy, rheology and the time varying macroscopic phase separation of the emulsions. For soybean oil and palm olein a rich diversity of emulsion microstructures and stabilities are realised. Five emulsion domains, each having a different microstructure and macroscopic stability have been identified within the composition space probed. For the lowest concentrations of emulsifier bridging flocculation is evident and emulsions are of low stability. Increasing Na-caseinate concentration leads to an increased stability and the existence of distinct individual oil droplets, visualised using cryo-scanning electron microscopy. Further increases in Na-caseinate concentration reduce emulsion stability due to depletion flocculation. Na-caseinate self-assembly is then initiated. At sufficiently high Na-caseinate and/or oil concentrations the continuous phase of the emulsion is a three-dimensional protein network and emulsion stability is again enhanced. At the limits of the emulsion composition space a gel-like paste is formed. The diversity of emulsion microstructure is reduced when tetradecane is the discrete phase. Na-caseinate self-assembly is limited and there is no evidence for formation of a protein network.

The association of hen eggs’ lysozyme with caseins was studied by using three casein substrates: (I) solutions of the various caseins, (II) artificially made casein micelles of various compositions and (III) caseins adsorbed onto soya-oil emulsion droplets. In solution, lysozyme associated most stro

The objective of this study was to monitor rennet-induced milk gel formation and mechanically induced gel syneresis in situ by low-field NMR. pH, temperature, and gel firmness at cutting time were varied in a factorial design. The new curve-fitting method Doubleslicing revealed that during coagulation two proton populations with distinct transverse relaxation times (T2,1=181, T2,2=465 ms) were present in fractions (f1=98.9%, f2=1.1%). Mechanical cutting of the gel in the NMR tube induced macrosyneresis, which led to the appearance of an additional proton population (T2,3=1500-2200 ms) identified as whey. On the basis of NMR quantification of whey water the syneresis rate was calculated and found to be significantly dependent on pH and temperature.

This chapter of the Encyclopedia of Dairy Science deals with the recent developments in the nomenclature, classification, structures, and associations of the major milk proteins: the caseins. Identification of the caseins continues to be based upon their primary structures. Significant findings re...

Polymorphism of goat milk alphas1-casein was determined and potential relations between genetic variants of this protein fraction and goat performance were evaluated. The investigations were performed on 598 goats assigned to of 4 breed groups (White improved 254 units, Coloured improved--124, White non-improved--146 and Coloured non-improved--74). For each goat, alphas1-casein polymorphism was determined in polyacrylamide gel by the PAGE-SDS method and percentage of milk alphas1-casein and gene frequency established. There was evaluated goat performance at successive lactations. In the goat population investigated, AA, AB, BB, AE, BE and EE alphas1-casein genotypes were identified. In all four breeds, alphas1-casein genotype EE clearly predominated (27.2-39.2%), recognized as "medium" and its share was higher in the groups of non-improved goats. It was conditioned by high frequency of gene E alphas1-casein (0.419-0.622). Generally, EE genotype percentage was higher in the non-improved goat groups. The improved goats, though, obtained higher productivity in each of the lactation studied. Analysis of relationships between alphas1-casein genetic variants and goats performance confirmed a significant influence on milk, protein and fat yields only in the Coloured improved goat group. There was revealed a more general tendency indicating a significant impact of "strong" alphas1-casein genotypes on a concentration of basic milk components, i.e. fat and protein, especially casein. In a group of goats producing milk of the highest casein content (over 2.4%) and protein (over 3.0%), the animals showing "strong" alphas1-casein variants dominated (85 and 70 %).

Casein micelle dispersions have been concentrated through osmotic stress and examined through rheological experiments. In conditions where the casein micelles are separated from each other, i.e., below random-close packing, the dispersions have exactly the flow and dynamic properties of the polydisperse hard-sphere fluid, demonstrating that the micelles interact only through excluded volume effects in this regime. These interactions cause the viscosity and the elastic modulus to increase by three orders of magnitude approaching the concentration of random-close packing estimated at Cmax≈178 g/l. Above Cmax, the dispersions progressively turn into "gels" (i.e., soft solids) as C increases, with elastic moduli G' that are nearly frequency independent. In this second regime, the micelles deform and/or deswell as C increases, and the resistance to deformation results from the formation of bonds between micelles combined with the intrinsic mechanical resistance of the micelles. The variation in G' with C is then very similar to that observed with concentrated emulsions where the resistance to deformation originates from a set of membranes that separate the droplets. As in the case of emulsions, the G' values at high frequency are also nearly identical to the osmotic pressures required to compress the casein dispersions. The rheology of sodium caseinate dispersions in which the caseins are not structured into micelles is also reported. Such dispersions have the behavior of associative polymer solutions at all the concentrations investigated, further confirming the importance of structure in determining the rheological properties of casein micelle systems.

-stranded gel network. Upon addition of high levels of calcium, cold gelation was promoted presumably through direct aggregation of casein micelles. Understanding cold gelation properties can facilitate potential use of RCM in cheese making.

Gelation is a significant operation in dairy processing. Protein gelation can be affected by several factors such as temperature, pH, or enzyme addition. Recently, the use of ultrasonication has been shown to have a significant impact on the formation of whey protein gels. In this work, the effect of ultrasonication on the gelation of casein systems was investigated. Gels were formed by the addition of 7.6 mm Tetra Sodium Pyro Phosphate (TSPP) to 5 wt% micellar casein (MC) solutions. Sonication at 20 KHz and 31 W for up to 30 min changed the surface hydrophobicity of the proteins, whereas surface charge was unaltered. Sonication before the addition of TSPP formed a firm gel with a fine protein network and low syneresis. Conversely, sonication after TSPP addition led to an inconsistent weak-gel-like structure with high syneresis. Gel strength in both cases increased significantly after short sonication times, while the viscoelastic properties were less affected. Overall, the results showed that ultrasonication can have a significant effect on the final gel properties of casein systems.

The effect of calcium chelators on physical changes of casein micelles in concentrated micellar casein solutions was investigated by measuring calcium-ion activity, viscosity and turbidity, and performing ultracentrifugation. The highest viscosities were measured on addition of sodium

The objective of this study was to analyze both the allergenicity and immunogenicity of cow's milk proteins. To this end, 80 milk-atopic patients were selected on the basis of the presence of cow's milk-specific IgE antibodies in serum and compatible clinical history. Fifteen patients allergic to other allergens and 10 nonatopic subjects were studied as controls. The specificity of serum IgG and IgE antibodies was determined by immunoblotting, employing both cow's milk and milk components, i.e., alpha- and beta-casein, beta-lactoglobulin, and alpha-lactalbumin separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The experiments showed that casein-specific IgE antibodies were present in all (80/80) sera examined; 10/80 showed reactivity to beta-lactoglobulin, and 5/80 showed reactivity to alpha-lactalbumin. None of the 25 negative control sera analyzed showed the presence of specific IgE antibodies against milk proteins. These results were similar to those corresponding to the detection, by the radioallergosorbent test, of IgE antibodies against the milk components coupled to paper disks. All sera from milk-atopic patients also showed IgE reactivity against a high-molecular-mass fraction that hardly enters the gel. This fraction, after separation by gel filtration and treatment with beta-mercaptoethanol and urea, was shown by SDS-PAGE analysis to be formed by casein monomers. All sera analyzed by immunoblotting reacted against the components corresponding to casein monomers. Inhibition of immunoblotting by adsorption with different milk components confirmed that those high-molecular-mass aggregates are formed by casein components. The results presented here strongly suggest that casein is the major allergenic component of cow's milk.

Casein micelles dispersions have been concentrated and equilibrated at different osmotic pressures using equilibrium dialysis. This technique measured an equation of state of the dispersions over a wide range of pressures and concentrations and at different ionic strengths. Three regimes were found. i), A dilute regime in which the osmotic pressure is proportional to the casein concentration. In this regime, the casein micelles are well separated and rarely interact, whereas the osmotic press...

The rheology of milk powder suspensions is investigated up to very high concentrations, where structural arrest occurs. The main component of the milk powder investigated is casein, so that the suspensions can be regarded as casein suspensions. Four concentration regimes are identified. For effective casein volume fractions less than 0.54 the concentration dependence of the zero-shear viscosity is similar to that of hard-sphere suspensions. However, due to the elastic deformation of the casei...

Full Text Available True lactose intolerance (symptoms stemming from lactose malabsorption is less common than is widely perceived, and should be viewed as just one potential cause of cows’ milk intolerance. There is increasing evidence that A1 beta-casein, a protein produced by a major proportion of European-origin cattle but not purebred Asian or African cattle, is also associated with cows’ milk intolerance. In humans, digestion of bovine A1 beta-casein, but not the alternative A2 beta-casein, releases beta-casomorphin-7, which activates μ-opioid receptors expressed throughout the gastrointestinal tract and body. Studies in rodents show that milk containing A1 beta-casein significantly increases gastrointestinal transit time, production of dipeptidyl peptidase-4 and the inflammatory marker myeloperoxidase compared with milk containing A2 beta-casein. Co-administration of the opioid receptor antagonist naloxone blocks the myeloperoxidase and gastrointestinal motility effects, indicating opioid signaling pathway involvement. In humans, a double-blind, randomized cross-over study showed that participants consuming A1 beta-casein type cows’ milk experienced statistically significantly higher Bristol stool values compared with those receiving A2 beta-casein milk. Additionally, a statistically significant positive association between abdominal pain and stool consistency was observed when participants consumed the A1 but not the A2 diet. Further studies of the role of A1 beta-casein in milk intolerance are needed.

True lactose intolerance (symptoms stemming from lactose malabsorption) is less common than is widely perceived, and should be viewed as just one potential cause of cows' milk intolerance. There is increasing evidence that A1 beta-casein, a protein produced by a major proportion of European-origin cattle but not purebred Asian or African cattle, is also associated with cows' milk intolerance. In humans, digestion of bovine A1 beta-casein, but not the alternative A2 beta-casein, releases beta-casomorphin-7, which activates μ-opioid receptors expressed throughout the gastrointestinal tract and body. Studies in rodents show that milk containing A1 beta-casein significantly increases gastrointestinal transit time, production of dipeptidyl peptidase-4 and the inflammatory marker myeloperoxidase compared with milk containing A2 beta-casein. Co-administration of the opioid receptor antagonist naloxone blocks the myeloperoxidase and gastrointestinal motility effects, indicating opioid signaling pathway involvement. In humans, a double-blind, randomized cross-over study showed that participants consuming A1 beta-casein type cows' milk experienced statistically significantly higher Bristol stool values compared with those receiving A2 beta-casein milk. Additionally, a statistically significant positive association between abdominal pain and stool consistency was observed when participants consumed the A1 but not the A2 diet. Further studies of the role of A1 beta-casein in milk intolerance are needed.

The analysis of casein polymorphisms in goat species is rather difficult, because of a large number of mutations at each locus, and the tight linkage involving the 4 casein genes. Three goat breeds from Northern Italy, Orobica, Verzasca, and Frisa, were analyzed at the casein complex by milk isoelectrofocusing and analyses at the DNA level to identify the majority of all known polymorphisms. The casein gene structure of the 3 local breeds at alpha(S1)-casein (CSN1S1), beta-casein (CSN2), alpha(S2)-casein (CSN1S2), and kappa-casein (CSN3) was compared with that of Camosciata, a more widely distributed breed. A new allele was identified and characterized at CSN2 gene, which seemed to be specific to the Frisa breed. It was named CSN2*E, and was characterized by a transversion TCT --> TAT responsible for the amino acid exchange Ser(166) --> Tyr(166) in the mature protein. The casein haplotype structure is highly different among breeds. A total of 26 haplotypes showed a frequency higher than 0.01 in at least 1 of the 4 breeds considered, with 12, 3, 5, and 19 haplotypes in Frisa, Orobica, Verzasca, and Camosciata breeds, respectively. Only 13 haplotypes occurred at a frequency higher than 0.05 in at least 1 breed. With the molecular knowledge of each locus, the ancestral haplotype coding for CSN1S1*B, CSN2*A, CSN1S2*A, and CSN3*B protein variants can be postulated. A protein evolutionary model considering the whole casein haplotype is proposed.

The development of advanced materials from biorenewable protein biopolymers requires the generation of more exogenous bonds to maintain the microstructure and durability in the final products. Casein is the main protein of milk, representing about 80% of the total protein. In the present investigation the casein protein was solubilized and/or emulsified in aqueous alkaline solutions, and 2D films and 3D matrices were produced. The effects of silane (3-aminopropyl triethoxy silane), DL-glyceraldehyde and glutaraldehyde on tensile properties and water swelling/absorption of 2D casein films and also the microstructure of the freeze-dried 3D matrices were analyzed. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis showed that there were no significant changes in the molecular weight (19-23.9 kDa) of the casein proteins on exposure to alkaline solutions of sodium hydroxide and silane. The casein films produced without glycerol plasticizer and with heat treatment (130 °C for 18 h) were fragile. However, the fragile films were transformed into ductile and tough materials on exposure to moisture (i.e., conditioned for one week at 50 ± 2% relative humidity and 22 ± 2 °C) and showed a maximum average tensile strength of 49-52 MPa and modulus of 1107-1391 MPa. The chemical cross-linkers (i.e., DL-glyceraldehyde and glutaraldehyde) improved the microstructure of glycerol plasticized casein protein, when analyzed under scanning electron microscope (SEM). Furthermore, these chemical cross-linking agents enhanced the mechanical properties and water resistant properties of casein films.

Gelation process of acid-induced caseingels was studied using response surface method (RSM). Ratio of casein to whey proteins, incubation and heating temperatures were independent variables. Final storage modulus (G') measured 200 min after the addition of glucono-δ-lactone and the gelation time i.e. the time at which G' of gels became greater than 1 Pa were the parameters studied. Incubation temperature strongly affected both parameters. The higher the incubation temperature, the lower was the G' and the shorter the gelation time. Increased heating temperature however, increased the G' but again shortened the gelation time. Increase in G' was attributed to the formation of disulphide cross-linkages between denatured whey proteins and casein chains; whilst the latter was legitimized by considering the higher isoelectric pH of whey proteins. Maximum response (G' = 268.93 Pa) was obtained at 2.7 % w/w, 25 °C and 90 °C for casein content, incubation and heating temperatures, respectively.

Full Text Available Milk is a complex biological fluid with high amount of proteins, lipid and minerals. The function of milk is to supply nutrients such as essential amino acids required for the growth of the newborn. In addition, due to the importance of casein and casein micelles for the functional behavior of dairy products, the nature and structure of casein micelles have been studied extensively. However, the exact structure of casein micelles is still under debate. Various models for casein micelle structure have been proposed. Most of the proposedmodels fall into three general categories, which are: coat-core, subunit (sub-micelles, and internal structure models. The coat-core models, proposed by Waugh and Nobel in 1965, Payens in 1966, Parry and Carroll in 1969, and Paquin and co-workers in 1987, describe the micelle as an aggregate of caseins with outer layer differing in composition form the interior, and the structure of the inner part is not accurately identified. The sub-micelle models, proposed by Morr in 1967, Slattery and Evard in 1973, Schmidt in 1980, Walstra in1984, and Ono and Obata in 1989, is considered to be composed of roughly spherical uniform subunits. The last models, the internal structure models, which were proposed by Rose in 1969, Garnier and Ribadeau- Dumas in 1970, Holt in 1992, and Horne in 1998, specify the mode of aggregation of the different caseins.

Casein micelles in milk are proteinaceous colloidal particles and are essential for the production of flocculated and gelled products such as yogurt, cheese, and ice-cream. The colloidal stability of casein micelles is described here by a calculation of the pair potential, containing the essential contributions of brush repulsion, electrostatic repulsion, and van der Waals attraction. The parameters required are taken from the literature. The results are expressed by the second osmotic virial coefficient and are quite consistent with experimental findings. It appears that the stability is mainly attributable to a steric layer of κ-casein, which can be described as a salted polyelectrolyte brush.

Full Text Available Casein, the main milk protein was determined by different assay methods: the gravimetric method, the method based on the neutralization of the NaOH excess used for the casein precipitate solving and the method based on the titration of the acetic acid used for the casein precipitation. The last method is the simplest one, with the fewer steps, and also with the lowest error degree. The results of the experiment revealed that the percentage of casein from the whole milk protein represents between 72.6–81.3% in experiment 1, between 73.6–81.3% in experiment 2 and between 74.3–81% in experiment 3.

We investigated complex coacervation of sodium caseinate/Astragalus rahensis (A.r) as a function of pH with light scattering, spectrophotometry, and viscosity measurements. Interestingly, sodium caseinate/A.r displayed five structural transitions; pH 7.00 to pH ∼5.40: no interaction occurred, pH ∼5.40 to pH ∼4.80: initiation of the formation of primary soluble complexes, pH ∼4.80 to ∼4.30: formation of interpolymer complexes, pH ∼4.30 to ∼4.02: optimum coacervation and pH ∼4.02 to ∼2.50: suppression of coacervation. In addition, rheological properties of sodium caseinate/A.r coacervates were studied at various pH values. A much higher storage modulus (G') than loss modulus (G″) for all sodium caseinate/A.r coacervates suggests the formation of highly interconnected gel-like network structures with mainly elastic behaviour. Moreover, sodium caseinate/A.r coacervates at all pH values exhibited a shear thinning behaviour across the entire shear rate range investigated. Effects of different species of gum tragacanth on the interactions with sodium caseinate have been scarcely studied. Our study showed that systems containing various species (A.r, soluble fraction of A.r and Astragalus gossypinus (A.g)) had different critical pH values and particle sizes during complex coacervation, which could be due to different ratio of soluble to insoluble fractions and uronic acid content of various species.

The internal structure of casein micelles was studied by calculating the small-angle neutron and X-ray scattering and static light scattering spectrum (SANS, SAXS, SLS) as a function of the scattering contrast and composition. We predicted experimental SANS, SAXS, SLS spectra self consistently using independently determined parameters for composition size, polydispersity, density and voluminosity. The internal structure of the casein micelles, i.e. how the various components are distributed within the casein micelle, was modeled according to three different models advocated in the literature; i.e. the classical sub-micelle model, the nanocluster model and the dual binding model. In this paper we present the essential features of these models and combine new and old experimental SANS, SAXS, SLS and DLS scattering data with new calculations that predict the spectra. Further evidence on micellar substructure was obtained by internally cross linking the casein micelles using transglutaminase, which led to casein nanogel particles. In contrast to native casein micelles, the nanogel particles were stable in 6 M urea and after sequestering the calcium using trisodium citrate. The changed scattering properties were again predicted self consistently. An important result is that the radius of gyration is independent of contrast, indicating that the mass distribution within a casein micelle is homogeneous. Experimental contrast is predicted quite well leading to a match point at a D{sub 2}O volume fraction of 0.41 ratio in SANS. Using SANS and SAXS model calculations it is concluded that only the nanocluster model is capable of accounting for the experimental scattering contrast variation data. All features and trends are predicted self consistently, among which the 'famous' shoulder at a wave vector value Q = 0.35 nm{sup -1}. In the nanocluster model, the casein micelle is considered as a (homogeneous) matrix of caseins in which the colloidal calcium phosphate (CCP

The changes in insulin concentration in bovine milk in the first period of lactation and its association with other milk proteins were studied. Highest concentration was found in the first milking (327 ng/ml). This concentration fell within the first 24 h postpartum to about 50% of its initial value. By d 3, the level was about 25%, and, on d 7, a stable concentration was reached at approximately 46 ng/ml (about 14% of its initial value). This concentration is about 100 times higher than that in serum, which suggests a specific mechanism of transfer from blood to milk. Colostral whey obtained by ultrafiltration or ultracentrifugation contains much less insulin than colostrum. When colostrum or milk was incubated with [125I]insulin and whey and casein fractions were separated by precipitation, it was observed that most insulin remained with the casein. However, when colostrum was incubated with [125I]insulin and subjected to gel filtration, most of the radioactivity corresponded to free insulin, indicating that insulin is associated with the precipitated casein but not with the casein micelles in solution.

The selective precipitation of whole casein from skimmed milk was achieved by the addition of calcium salt under acidic pH. The effects of pH, centrifugal force and final concentration of CaCl2 on the separation of casein were studied by measuring the purity of final products using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The results showed that casein with the highest purity could be obtained with the pH of 4.3, the centrifugal force of 10 400 g and the final concentration of CaCl2 of 60 mmol/L. The casein was processed with DEAE anion exchange chromatography and three peaks were obtained. Then the third peak (peak III) was identified with Western-Blot method and matrix-assisted laser desorption/ionization time of flight/time of flight mass spectrometry (MALDI-TOF/TOF MS). The identification of Western-Blot showed that peak III can combine with the specificity of human milk beta-casein antibody, and it is proved to be human milk beta-casein. The fingerprints of peak III were nalyzed by Mascot searching, and the sequence coverage was 50%, further supporting it is human milk beta-casein. In conclusion, an effective method to obtain human milk beta-casein from milk samples through DEAE anion exchange chromatography was established, and it is suitable for the proteomics research requirements of the beta-casein from human milk.

The general aim of the work described in this thesis was to investigate structure-rheologyrelations for dairy related products, focusing on model systems containing sodium caseinate. The acid inducedgelationof sodium caseinate, of sodium caseinate

Casein and whey differ in amino acid composition and in the rate of absorption; however, the absorption rate of casein can be increased to mimic that of whey by exogenous hydrolysis. The objective of the present study was to compare the effects of hydrolysed casein (HC), intact casein (IC......) and intact whey (IW) on energy expenditure (EE) and appetite regulation, and thereby to investigate the influence of amino acid composition and the rate of absorption. In the present randomised cross-over study, twenty-four overweight and moderately obese young men and women consumed three isoenergetic...... dietary treatments that varied in protein source. The study was conducted in a respiration chamber, where EE, substrate oxidation and subjective appetite were measured over 24 h at three independent visits. Moreover, blood and urine samples were collected from the participants. The results showed...

The average casein micelle size varies widely between milk samples of individual cows. The factors that cause this variation in size are not known but could provide more insight into casein micelle structure and into the physiology of casein micelle formation. The objective of this research was ther

Only recently, researchers have started experimenting with 3D printing of foods. The aim of this study was to investigate 3D printed objects from sodium caseinate dispersions, exhibiting reversible gelation behaviour. Gelation and dispensing behaviour were explored and structures of different pro

Heating of milk is an essential step in the processing of various dairy products, like for example yoghurt. A major consequence of the heat treatment is the denaturation of whey proteins, which either associate with the casein micelle or form soluble whey protein aggregates. By combination of enzyma

The aim of this work was to improve an existing method to separate and quantify the 4 major caseins from milk samples (i.e., containing whey proteins) using ion-exchange chromatography. The separation process was carried out using a mini-preparative cation exchange column (1 or 5mL of column volume), using urea acetate as elution buffer at pH 3.5 with a NaCl gradient. All 4 major caseins were separated, and the purity of each peak was assessed using sodium dodecyl sulfate-PAGE. Purified casein fractions were also added to raw milk to confirm their elution volumes. The quantification was carried out using purified caseins in buffer as well as added directly to fresh skim milk. This method can also be employed to determine the decrease in kappa-casein and the release of the casein-macropeptide during enzymatic hydrolysis using rennet. In this case, the main advantage of using this method is the lack of organic solvents compared with the conventional method for separation of macropeptide (using reversed phase HPLC).

Full Text Available Objectives: This study aimed to assess the effect of applying casein phosphopeptide–amorphous calcium phosphate (CPP-ACP paste, casein phosphopeptide–amorphous calcium phosphate fluoride (CPP-ACPF paste and sodium fluoride gel on surface microhardness of enamel after microabrasion.Materials and Methods: Thirty freshly extracted human premolars were selected. All samples were subjected to hardness indentations made with the Vickers hardness machine and the average value was recorded as the initial surface microhardness. The specimens were then randomly divided into three groups (n=10 of CPP-ACPF, fluoride and CPP-ACP. The teeth were micro-abraded with Opalustre. Microhardness test was performed to assess the post-abrasion hardness. Three remineralization modalities were performed on samples of each group. The enamel surface microhardness measurements were performed. To compare the difference between groups, the rehardening and softening values were defined. One-way ANOVA and Tukey’s post hoc test at a significance level of 5% were used for statistical analysis.Results: The mean microhardness value (MMV had a significant decrease after microabrasion from baseline. The MMV had a significant increase after remineralization in all groups. The MMV of CPP-ACPF group was significantly more than that of fluoride group (P=0.027. The rehardening value of fluoride group was significantly more than that of other groups (P<0.001.Conclusion: All the remineralizing agents were effective for rehardening the enamel after microabrasion. The CPP-ACP and CPP-ACPF pastes are effective, but to a lesser extent than neutral sodium fluoride gel in remineralizing enamel surface. Incorporation of fluoride to CPP-ACP formulation does not provide any additional remineralizing potential.Keywords: Casein phosphopeptide-amorphous calcium phosphate nanocomplex; Enamel Microabrasion; Hardness; Sodium Fluoride

In this study, the effects of texture modification and aroma-induced sweetness enhancement were systematically investigated in apple-flavored semi-solid Na-caseinategels. Gels containing apple juice as a basic flavor were developed differing in stiffness, brittleness and serum release (texture modi

The aim of this work was to investigate the impact on the texture properties of emulsion-filled gels when saturated solid fat is replaced by unsaturated liquid oil. Whey protein aggregate, gelatin and micellar casein, were chosen to form different types of gel matrices and the fat hardness was varie

β-Casein was the main component of human milk casein, but the content of β-casein in the bovine milk was less. The difference in β-casein content of the two samples was one of the reasons why human milk is more digestible than bovine milk. Studying the differences of structure and function in human and bovine milk β-casein can help us develop a new human milk simulated infant formula which will be more suitable for the infant gut. The UV spectrophotometer was used to study the solubility, sulfhydryl and emulsification of human milk β-casein and bovine milk β-casein, Fluorescence spectroscopy and the infrared spectroscopy were used to study the structural characteristics of human milk β-casein and bovine milk β-casein. The two samples shared a similar isoelectric point (pH 4.0~5.0), the solubility of human milk β-casein (10.83%) was lower than which in bovine milk β-casein (11.83%) near the pI, while it was higher when it deviated the pI. The emulsion ability (110~140 m2 · g(-1)) of human milk β-casein was higher than that in bovine milk β-casein (70~130 m2 · g(-1)) and surface sulfhydryl group (SH) of two kinds of milk protein were similar [(18.47±0.08) μmol · g(-1) and (18.67±0.17) μmol · g(-1)]. The total sulfhydryl group [(47.46±0.23) μmol · g(-1)] in bovine milk β-casein was more than that in human milk β-casein [(26.17±0.12) μmol · g(-1)]. Functional groups in two samples were similar and they both contained beta sheet, human milk β-casein had less H-bond and internal hydrophobic than bovine milk β-casein. The results showed that the two samples had similar functional groups, while human milk β-casein had much less secondary structure such as α-helix and β-sheet, a looser tertiary structure and a better interfacial activity.

Full Text Available Point mutations in exon IV of the bovine κ-casein (CSN3 gene determine two allelic variants, A and B. These variants were distinguished by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP analysis in the indigenous Sahiwal and Tharparkar cattle breeds. DNA samples (252 Sahiwal and 56 Tharparkar were analyzed for allelic variants of the CSN3 gene. Polymorphism was detected by digestion of PCR-amplified products with HindIII, HhaI and HaeIII restriction enzymes, followed by separation on 3% agarose gels, and resolved by ethidium bromide staining. Allele A of the κ-casein gene occurred at a higher frequency than allele B, in both Sahiwal and Tharparkar breeds. The genotypic frequencies of AA, AB, and BB in the Sahiwal and Tharparkar breeds were 0.758, 0.230 and 0.012, and 0.0.732, 0.250 and 0.018, respectively. The frequencies of alleles A and B in the Sahiwal and Tharparkar breeds were 0.873 and 0.127, and 0.857 and 0.143, respectively. Genotype BB of the kappa-casein gene had more influence on the monthly milk yield, 305-days milk yield, monthly solids-not-fat (SNF yield, and monthly protein yield, in the Sahiwal cattle.

This research is mainly focusing on one-step biosynthesis of graphene from graphene oxide and its stabilization using naturally occurring milk protein, casein. The synthesis of casein reduced graphene oxide (CRGO) was completed within 7h under reflux at 90°C with the formation of few layered fine graphene nanosheets. UV-Vis, XRD, XPS analysis data revealed the reduction process of the graphene oxide. Results of FT-IR, HPLC and TEM analysis have shown that the ensuing material consists of graphene decorated with casein molecules. Aspartic acid and glutamic acid residue present in casein molecules are responsible for the reduction of graphene oxide.

This research is mainly focusing on one-step biosynthesis of graphene from graphene oxide and its stabilization using naturally occurring milk protein, casein. The synthesis of casein reduced graphene oxide (CRGO) was completed within 7 h under reflux at 90 °C with the formation of few layered fine graphene nanosheets. UV-Vis, XRD, XPS analysis data revealed the reduction process of the graphene oxide. Results of FT-IR, HPLC and TEM analysis have shown that the ensuing material consists of graphene decorated with casein molecules. Aspartic acid and glutamic acid residue present in casein molecules are responsible for the reduction of graphene oxide.

Solidification, emulsification and application of shear were combined to induce diversity and heterogeneity in the micro- and macrostructure of concentrated caseinate-based food matrices containing a dispersed fat phase. The products were evaluated with selected parameters from small-scale and large

Sialic acid is a carbohydrate moiety of k-casein glycomacropeptide (GMP), which is a 64 amino acid residue C-terminal sialylated phosphorylated glycopeptide released from k-casein by the action of chymosin during cheese making. GMP lacks aromatic amino acids including phenylalanine, tyrosine, and tryptophan. Because of its unique amino acid composition and various biological activities, GMP is thought to be a potential ingredient for dietetic foods (e.g., a food for PKU patients) and pharmaceuticals. Thus, increased attention has been given to the development of techniques to purify GMP. In this review, techniques of GMP purification described in patents and scientific research papers were introduced. A sialic acid assay is the important method to track GMP isolation and purification processes, for which the thiobarbituric acid reaction with 1-propanol as a chromophore extracting solvent is an inexpensive, practical and specific technique. Sephacryl S-200 gel filtration chromatography, cellulose acetate electrophoresis, and sodium dodecyl sulfate polyacrylamide gel electrophoresis are the major techniques to identify sialic acid specific to GMP. Sephacryl S-200 chromatography and cellulose acetate electrophoresis are also used to detect GMP sialic acid in whey pearmeate and whey added commercial margarine samples. Future research includes development of an economical industrial scale method to produce high purity GMP.

This book chapter deals with the molecular architecture of the major milk proteins, the caseins. Earlier theories of casein structure classified them as random coils; i.e., flexible without definite structural elements. Recent advances in the field of protein chemistry have significantly enhanced ...

Casein phosphorylation is a posttranslational modification catalyzed by kinase enzymes that attach phosphate groups to specific AA in the protein sequence. This modification is one of the key factors responsible for the stabilization of calcium phosphate nanoclusters in casein micelles and for the i

Full Text Available Because more than 60% of milk produced in Italy is transformed into cheese, milk economical value strongly depends on cheese yield. Among the factors that influence cheese yield, milk casein and fat content plays a major role: when milk is converted into Grana Padano and Parmigiano reggiano, three grams of seasoned cheese are produced from one gram of milk casein.....

Melanoidins are the final, brown, high molecular weight products of the Maillard reaction. The aim of the present study was to determine the average molar extinction coefficients of melanoidins formed in heated glucose-casein and fructose-casein systems. The value of the extinction coefficient can b

Casein micelles in skim milk were either untreated (untreated milk) or were cross-linked using transglutaminase (TGA-milk). Added lactoferrin (LF) bound to the casein micelles and followed Langmuir adsorption isotherms. The adsorption level was the same in both milks and decreased the micellar zeta

We explore the properties of casein/xanthan mixtures for xanthan concentrations beyond those inducing phase separation. Previous work has successfully described the onset of demixing by depletion theory in the protein limit, where the xanthan polysaccharides, the polymers, are larger than the casein

A new process for restructured meat and fish has been introduced to the market recently. Its main compound is casein, and it may therefore endanger patients with a milk allergy. Chemicals/CAS: Anti-Inflammatory Agents; Caseins; Lactoglobulins; Methylprednisolone, 83-43-2

Melanoidins are the final, brown, high molecular weight products of the Maillard reaction. The aim of the present study was to determine the average molar extinction coefficients of melanoidins formed in heated glucose-casein and fructose-casein systems. The value of the extinction coefficient can

Initially the resistance to digestion of two cow's milk allergens, beta-casein, and beta-lactoglobulin (beta-Lg), was compared using a "high-protease assay" and a "low-protease assay" in a single laboratory. The low-protease assay represents an alternative standardised protocol mimicking conditions...... found in the gastrointestinal tract. For the high-protease assay, both proteins were incubated with either pepsin or pancreatin and digestion monitored by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and reverse phase-high performance liquid chromatography. The low-protease assay involved...... gastroduodenal digestion in the presence or absence of phosphatidylcholine (PC). Both beta-casein and beta-Lg were susceptible to hydrolysis by pepsin and pancreatin in the high-protease assay. In contrast, the kinetics of beta-casein digestion in the low-protease assay were slower, beta-Lg being pepsin...

Heat-induced aggregation and gelation of aqueous solutions of whey protein isolate (WPI) in the presence of sodium caseinate (SC) and CaCl2 was studied at pH 6.6. The effect of adding SC (0-100 g/L) on the structure of the aggregates and the gels was investigated by light scattering and confocal laser scanning microscopy at different CaCl2 concentration ([CaCl2] = 0-30 mM). The gelation process was studied by oscillatory shear rheology. At the whey protein concentrations studied here (34 and 60 g/L), no gels were formed in the absence of CaCl2 and SC. However, WPI solutions gelled above a critical CaCl2 concentration that increased with increasing SC concentration. In the absence of CaCl2, WPI gels were formed only above a critical SC concentration. The critical SC concentration needed to induce WPI gelation decreased weakly when CaCl2 was added. In an intermediate range of CaCl2 concentrations, gels were formed both at low and high SC concentrations, but not at intermediate SC concentrations. Finally, at high CaCl2 concentrations gels were formed at all SC concentrations. The gelation rate and the gel structure of the gels formed at low and high casein concentrations were very different. The effect of SC on the thermal gelation of WPI was interpreted by competition for Ca(2+), a chaperon effect, and microphase separation.

Nucleolin (C23), a 110 kDa phosphoprotein, which is mainly found in the nucleolus has been shown to be a physiological substrate for casein kinase II (CKII). Nucleolin was identified and characterized by immunodetection using an anti-nucleolin antibody. Phosphopeptide patterns from nucleolin...... phosphorylated by purified casein kinase II and of phosphorylated nucleolin which had been isolated from tumor cells grown in the presence of [32P]-o-phosphate, were identical. The partial tryptic digest revealed nine phosphopeptides. Nucleolin isolated from Krebs II mouse ascites cells was phosphorylated...... by purified casein kinase II with about two moles phosphate per one mole of nucleolin....

The aim of this work was to develop an alginate-casein composite microsphere as a bioactive vehicle for oral administration of nutrients by a simple extrusion dripping method. Riboflavin was selected as a model drug, and the microencapsulation efficiency was raised to 97.94%after optimizing the preparation conditions by response surface methodology. In vitro release studies showed that riboflavin was released completely from alginate-casein microspheres in simulated intestinal fluids. Meanwhile, the morphology, structure and interaction between alginate and casein were characterized by scanning electron microscopy (SEM) and Fourier transform infrared (FTIR) spectra.

Full Text Available Mechanical properties have been widely correlated with textural characteristics to determine the interactions during the process formation of dairy gel. These interactions are strongly affected by process conditions and system composition. In the present study, the rheological of acid-induced protein dairy gels with (2(7-3 and without (2(6-2 sucrose and subjected to small and large deformations were studied using an experimental design. The independent variables were the sodium caseinate, whey protein concentrate (WPC, carrageenan and sucrose concentrations as well as stirring speed and heat treatment time and temperature. Mechanical deformation tests were performed at 0.1, 1, 5, and 9 mm/s up to 80% of initial height. A heavy dependence of rupture stress on increasing crosshead speed and the formation of harder gels with the addition of sucrose were observed. Moreover the elastic and viscous moduli, obtained by fitting the Maxwell model to stress relaxation data, increased with increasing addition of sucrose. These results can be explained by preferential hydration of the casein with sucrose, causing an induction of casein-polysaccharide and casein-casein interactions.

The main objective of this study was to determine potential application of a serine proteinase derived from Asian pumpkin for obtaining biologically active peptides from casein. The course of casein hydrolysis by three doses of the enzyme (50, 150, 300 U/mg of protein) was monitored for 24 hours by the determinations of: hydrolysis degree DH (%), free amino group content (μmole Gly/g), RP HPLC peptide profiles and by polyacrylamide gel electrophoresis. In all hydrolyzates analyzed antioxidant activities were determined using three tests: the ability to reduce iron ions in FRAP test, the ability to scavenge free radicals in DPPH test, and Fe(2+) chelating activity. The antimicrobial activity of obtained peptide fractions was determined as the ability to inhibit the growth of Escherichia coli, Bacillus cereus and Pseudomonas fluorescens in a diffusion plate test. The deepest degradation, expressed as the DH [%] and the free amino group content (67% and 7528 µmole Gly/mg, respectively), was noted in samples hydrolyzed with 300 U/ml of enzyme for 24 hours, while in other samples the determined values were about three and two times lower. The results were in agreement with the peptide profiles obtained by RP HPLC. The highest antioxidative activities determined in all tests were seen for the casein hydrolysate obtained with 300 U/mg protein of serine proteinase after 24 h of reaction (2.15 µM Trolox/mg, 96.15 µg Fe(3+)/mg, 814.97 µg Fe(2+)/mg). Antimicrobial activity was presented in three preparations. In other samples no antimicrobial activity was detected.

Full Text Available Application of sodium caseinate as a functional additive in manufacturing processes requires production of its concentrated aqueous solutions which, in industrial conditions, presents a number of difficulties. In order to develop an effective and optimal industrial process of mixing – manufacturing a concentrated solution of sodium caseinate, it is essential to know rheological properties in a definite range of concentrations changing in the course of the dissolving process. The material for investigations was typical commercial sodium caseinate in the form of dry powder manufactured in Poland from acid casein using the method of extrusion. The objective of the undertaken empirical studies was the assessment of the impact of the concentration on rheological properties of sodium caseinate concentrates. Investigations were carried out for five concentrates manufactured in a mixer equipped in a mechanical agitator at concentrations ranging X (% Î (2.5¸12.5 and changing mass proportions of sodium caseinate in the aqueous solution as follows: GS/G (kgS·kg-1 = 0.025. On the basis of the obtained research results, classical flow curves were plotted for individual concentrates. The determined values of viscosity and density of the examined solutions were correlated in the form of h = f(GS/G and r = f(GS/G dependencies which were used during the determination of classical characteristics of mixing forces essential for the assessment of energetic expenditures required to manufacture concentrates in a mixer equipped in a mechanical agitator. The density of the examined concentrates increased in a way directly proportional, while the dynamic viscosity coefficient increased exponentially together with the increase of sodium caseinate concentration. Sodium caseinate concentrates exhibited Newtonian character in the examined range of concentrations.

We have optimized the polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) technique to screen the most frequent variants (A1, A2, A3, and B) of the bovine beta-casein gene. Five partly overlapping PCR products (233, 234, 265, 466, and 498 bp) of Exon VII of the beta-casein gene that encompass the target point mutations were heat-denatured, separated on nondenaturing polyacrylamide gels, and silver-stained. Simultaneous detection of all variants in reference samples of known genotypes (A1A2, A2A2, A1A3, A1B, and A2B) was best achieved on 17% polyacrylamide (100:1 acrylamide:bis-acrylamide ratio) gels with the PCR product of 234 bp. These results were confirmed by sequencing the allele-specific SSCP bands directly excised from polyacrylamide gels. A population of 65 anonymous samples belonging to various breeds was then analyzed twice, without discrepancies in a blind trial. Routine beta-casein genotyping using PCR-SSCP is proposed as a cost-effective, fast, and sensitive technique.

Nine mutants of human casein kinase-2 beta subunit have been created and assayed for their ability to assemble with the catalytic alpha subunit to give, at a 1:1 molar ratio, a fully competent CK-2 holoenzyme as judged by the following criteria: 1) the generation of an active heterotetrameric form...... of CK-2 exhibiting the expected sedimentation coefficient and 2) the enhancement of catalytic activity of CK-2 alpha. Extended deletions of 71 and 44 residues from the C-terminal end, but not a 7 residue deletion (including the cdc2 phosphorylation site) prevent both reconstitution of the holoenzyme and......-70 sequence give rise to mutants that still assemble with the alpha subunit to give a tetrameric holoenzyme. However, in the case of the mutants A57,59, A63,64, A59-61,63,64 in vitro assembly with the CK-2 alpha subunit was not complete. There were also intermediate complexes, free alpha-subunit and beta...

Full text: The most frequent destabilization mechanisms in Sodium Caseinate (NaCas) emulsions are creaming and flocculation. Coarse or fine emulsions with low protein con- tent destabilize mainly by creaming. If migration mechanism is suppressed, flocculation may become the main mechanism of destabilization. Small Angle X-Ray Scattering (SAXS) technique was applied to investigate sodium caseinate micelles structure in different environments. As many natural products, Sodium Caseinate samples have large polydisperse size distribution. The experimental data was analyzed using advanced modeling approaches. The Form Factor for the Caseinate micelle subunits was described by an ellipsoidal core shell model and the structure factor was split into two contributions, one corresponding to the particle-particle interactions and another one for the long range correlation of the subunits in the supramolecular structure. For the first term the hard sphere structure factor using the Percus-Yevick approximation for closure relation was used and for the second term a fractal model was applied. Three concentrations of sodium Caseinate (2, 5 and 7.5 %wt.) were measured in pure water, sugar solutions (20 %wt.) and in three different lipid phase emulsions containing 10 %wt. sunflower seed, olive and fish oils. Data analysis provided an average casein subunit radius of 4 nm, an average distance between the subunits of around 20nm and a fractal dimension value of around 3 for all samples. As indicated by the values of the correlation lengths for the set of studied samples, the casein aggregation is strongly affected by simple sugar additions and it is enhanced by emulsion droplets hydrophobic interaction. As will be presented, these nanoscale structural results provided by scattering experiments is consistent with macroscopic results obtained from several techniques, providing a new understanding of NaCas emulsions. (author)

Highly concentrated micellar casein concentrate (HC-MCC), a potential ingredient of protein-fortified food, is a gel at cold temperature. It contains ~17 to 21% casein, with most serum proteins and lactose removed by microfiltration and diafiltration, and it is then further concentrated using vacuum evaporation. The HC-MCC can be stored frozen, and our objective was to determine the conditions needed to obtain complete solubility of thawed HC-MCC in water and to understand its gelation upon cooling. Dispersibility (ability to pass through a 250-µm mesh sieve), suspendability (percentage of protein not sedimented at 80 × g within 5min), and solubility (percentage of protein not sedimented at 20,000 × g within 5min) were measured at 4, 12, or 20°C after various mixing conditions. Gelation upon cooling from 50 to 5°C was monitored based on storage (G') and loss (G'') moduli. The gelled HC-MCC was also examined by transmission electron microscopy. Thawed HC-MCC was added to water to reach a protein concentration of 3% and mixed using high shear (7,500rpm) for 1min or low shear (800rpm) for 30min at 4, 12, 20, or 50°C and at pH 6.4 to 7.2. The HC-MCC completely dispersed at 50°C, or at ≤20°C followed by overnight storage at 4°C. Suspendability at 50°C was ~90% whereas mixing at ≤20°C followed by overnight storage resulted in only ~57% suspendability. Solubility followed a similar trend with ~83% at 50°C and only ~29% at ≤20°C. Mixing HC-MCC with 60mM trisodium citrate increased dispersibility to 99% and suspendability and solubility to 81% at 20°C. Cold-gelling temperature, defined as the temperature at which G'=G'' when cooling from 50 to 5°C, was positively correlated with protein level in HC-MCC. Gelation occurred at 38, 28, and 7°C with 23, 20, and 17% of protein, respectively. Gelation was reversible upon heating, although after a second cooling cycle the HC-MCC gel had lower G'. In micrographs of gelled HC-MCC, the casein micelles were

Full Text Available Casein glycomacropeptide (GMP is known to promote the in vitro growth of Bifidobacteria and Lactobacilli. In this paper, we used conven- tional culture techniques and fluorescent in situ hybridization (FISH techniques to investigate the effect of casein GMP on mice fecal microbiota. The population structure of the intestinal microbiota, including Lactobacillus, Bifidobacteria, Enterococcus, coliforms and Enterobacteriaceae, was tested and compared. After consecutive administration of casein GMP for 15 days, numbers of Lactobacillus and Bifidobacteria increased significantly (P<0.01, numbers of Enterobacteriaceae and Coliforms decreased significantly (P<0.05 while no significant changes were observed for Enterococcus. The detection limits of FISH technique were significantly lower (P<0.01 than the traditional culture method. These results suggested that consumption of casein GMP had a prebiotic effect on male BALB/c mice. Casein GMP helped establish a healthier intestinal microbiota. Additionally, FISH was proved to be a rapid and relatively low-cost detection method that can be used to further our understanding of human intestinal microbiota.

Full Text Available Abstract Microtubules and associated motor proteins such as kinesin are envisioned for applications such as bioseparation and molecular sorting to powering hybrid synthetic mechanical devices. One of the challenges in realizing such systems is retaining motor functionality on device surfaces. Kinesin motors adsorbed onto glass surfaces lose their functionality or ability to interact with microtubules if not adsorbed with other supporting proteins. Casein, a milk protein, is commonly used in microtubule motility assays to preserve kinesin functionality. However, the mechanism responsible for this preservation of motor function is unknown. To study casein and kinesin interaction, a series of microtubule motility assays were performed where whole milk casein, or its αs1 and αs2, β or κ subunits, were introduced or omitted at various steps of the motility assay. In addition, a series of epifluorescence and total internal reflection microscopy (TIRF experiments were conducted where fluorescently labeled casein was introduced at various steps of the motility assay to assess casein-casein and casein-glass binding dynamics. From these experiments it is concluded that casein forms a bi-layer which supports the operation of kinesin. The first tightly bound layer of casein mainly performs the function of anchoring the kinesin while the second more loosely bound layer of casein positions the head domain of the kinesin to more optimally interact with microtubules. Studies on individual casein subunits indicate that β casein was most effective in supporting kinesin functionality while κ casein was found to be least effective.

Addition of calcium chloride to milk has positive effects on cheese-making because it decreases coagulation time, creates firmer gels, and increases curd yield. Although addition of calcium chloride is a widely used industrial practice, the effect of soluble calcium on the preliminary stages of gelation is not fully understood. In addition, it is not known whether the manner of addition and equilibration of the soluble calcium would affect the rennetability of the casein micelles. Therefore, the aim of this paper was to study the details of the coagulation behavior of casein micelles in the presence of additional calcium, and to elucidate whether the manner in which this cation is added (directly as calcium chloride or by gradual exchange through dialysis) affects the functionality of the micelles. Calcium was added as CaCl(2) (1 mM final added concentration) directly to skim milk or indirectly using dialysis against 50 volumes of milk. Additional soluble calcium did not affect the primary phase of the renneting reaction, as demonstrated by the analysis of the casein macropeptide (CMP) released in solution; however, it shortened the coagulation time of the micelles and increased the firmness of the gel. The turbidity parameter of samples with or without calcium showed that similar amounts of CMP were needed for particle interactions to commence. However, the amount of CMP released at the point of gelation, as indicated by rheology, was lesser for samples with added calcium, which can be attributed to a greater extent of calcium bridging on the surface or between micelles. The results also showed that the manner in which calcium was presented to the micelles did not influence the mechanism of gelation.

The size distribution and protein composition of casein micelles in the milk of Holstein-Friesian cows was determined as a function of stage and number of lactations. Protein composition did not vary significantly between the milks of different cows or as a function of lactation stage. Differences in the size and polydispersity of the casein micelles were observed between the milks of different cows, but not as a function of stage of milking or stage of lactation and not even over successive lactations periods. Modal radii varied from 55 to 70 nm, whereas hydrodynamic radii at a scattering angle of 73° (Q² = 350 μm⁻²) varied from 77 to 115 nm and polydispersity varied from 0.27 to 0.41, in a log-normal distribution. Casein micelle size in the milks of individual cows was not correlated with age, milk production, or lactation stage of the cows or fat or protein content of the milk.

This paper deals with the expression and distribution of casein kinase 2 (CK-2) subunits in mouse embryos at different developmental stages. Expression was investigated at the mRNA level of CK-2 alpha- and beta-subunits by in situ hybridization and distribution at the protein level by immunohisto......This paper deals with the expression and distribution of casein kinase 2 (CK-2) subunits in mouse embryos at different developmental stages. Expression was investigated at the mRNA level of CK-2 alpha- and beta-subunits by in situ hybridization and distribution at the protein level...

Full Text Available An amperometric biosensor for the quantification of food allergens based on an inhibitory immunoassay is presented. As a proof of concept, the experimental conditions were optimized for the detection of β-casein in the 0-10 ppm range. Eight electro­che­mi­cal cells were integrated into a small-sized portable potentiostat controlled by a smart­phone via Bluetooth communication. The determination of β-casein in eight different samples can be measured with the electrochemical biosensor, which has the potential to be modified for the detection of multiple allergens.

Heterologous conjugates of wheat arabinoxylan and β-casein were prepared via enzymatic cross-linking, using sequential addition of the arabinoxylan to a mixture of β-casein, peroxidase, and hydrogen peroxide. The maximal formation of adducts between the β-casein and the feruloylated arabinoxylan was

Full Text Available Bioactive peptide from casein play a very important role in biological functionalities such as antioxidant and antimicrobial activities. Casein is the main protein that derived from goat milk which consists of alpha (α, beta (β and kappa (κ casein. Dietary protein such as casein from animal can provide rich source of bioactive peptide. However, the macromolecular protein such as cow milk can cause allergic response to certain individuals. On the other hand, goat milk have been known for its hypoallergenic and therapeutic properties in human nutrition and health. The purpose of this study is to extract casein from local breed goat milk and identify the molecular characterization of casein for nutraceutical value. The casein was successfully extracted using extraction method. Extraction is a common technique used to separate a desired substance when it is mixed with other components. The average percentage of casein obtained was 24.25%. Then, the casein was analysed by running it in the SDS-Page. The major fraction is β-casein and the minor is α-casein that can be seen between 20kDa and 30kDa respectively. There is no contaminated protein appear in the purified α-amylase. The result obtained in this study indicates that isolated casein from Malaysian goat milk was pure and can be used as bioactive peptide for nutraceutical value.

A Ca(2+)-stimulated protein kinase (PK-I), active with dephosphorylated casein as exogenous substrate, was purified from ripening mango fruit. The purification procedure involved 30-70% ammonium sulphate fractionation and sequential anion exchange-, affinity-, hydrophobic interaction- and gel filtration chromatography. PK-I was purified ca. 40-fold with an overall yield of inhibition studies with ADP as product inhibitor best fit an ordered bi-bi kinetic mechanism with the Mg(2+)-ATP complex binding first to the enzyme followed by binding of the protein substrate. The K(m)ATP and K(m)casein of PK-I were 9 microM and 0.26 mg ml-1, respectively. The KiADP of PK-I was 9 microM.

The aggregation of casein micelles (CMs) induced by milk-clotting enzymes is a process of fundamental importance in the dairy industry for cheese production; however, it is not well characterized on the nanoscale. Here we enabled the monitoring of the kinetics of aggregation between single CMs (30...

ß-Casein, which is present in the form of micelles at atmospheric pressure, has been hydrolyzed during pressure treatment to improve the accessibility of the protein. Two proteolytic enzymes with different specificities were used. Trypsin was aimed at mainly hydrolyzing hydrophilic segments of ß-cas

The reaction of malondialdehyde with casein was studied in aqueous solution to evaluate the impact of this lipid oxidation product on food protein modification. By using multiresponse modeling, a kinetic model was developed for this reaction. The influence of temperature and pH on protein browning a

In the Mediterranean region, goat milk production is an important economic activity. In the present study, 4 casein genes were genotyped in 5 Sicilian goat breeds to 1) identify casein haplotypes present in the Argentata dell'Etna, Girgentana, Messinese, Derivata di Siria, and Maltese goat breeds; and 2) describe the structure of the Sicilian goat breeds based on casein haplotypes and allele frequencies. In a sample of 540 dairy goats, 67 different haplotypes with frequency >or=0.01 and 27 with frequency >or=0.03 were observed. The most common CSN1S1-CSN2-CSN1S2-CSN3 haplotype for Derivata di Siria and Maltese was FCFB (0.17 and 0.22, respectively), whereas for Argentata dell'Etna, Girgentana and Messinese was ACAB (0.06, 0.23, and 0.10, respectively). According to the haplotype reconstruction, Argentata dell'Etna, Girgentana, and Messinese breeds presented the most favorable haplotype for cheese production, because the casein concentration in milk of these breeds might be greater than that in Derivata di Siria and Maltese breeds. Based on a cluster analysis, the breeds formed 2 main groups: Derivata di Siria, and Maltese in one group, and Argentata dell'Etna and Messinese in the other; the Girgentana breed was between these groups but closer to the latter.

Casein kinase II (CKII) activity is enhanced as much as 2-3 fold in established and 4-5-fold in transformed human cell lines when compared to that of fibroblasts and primary human tumour cell cultures where CKII activity never exceeded a basic level. The high activity of CKII in transformed cells...

In this paper, the structure, characteristics, modification methods and application of casein are summarized. It is hoped that this can be helpful to related researchers.% 综述了酪素的结构、特性、改性方法和用途，希望能作为相关科技工作者的参考。

The size distribution and protein composition of casein micelles in the milk of Holstein-Friesian cows was determined as a function of stage and number of lactations. Protein composition did not vary significantly between the milks of different cows or as a function of lactation stage. Differences i

The physicochemical properties of acidified milk gels after the addition of cocoa flavanols were studied. As the flavanol level increased (from 0 to 2.5 mg/g), syneresis and gel elasticity (tan δ) were found to significantly increase and decrease, respectively. Flavanol addition reduced the stress at fracture, with no changes in fracture strain, suggesting that the bond type (i.e., covalent vs noncovalent) was the underlying factor explaining the ease of fracture. Gels made from recombined milks containing the casein fraction of heated milk and the serum of heated flavanol/milk mixtures showed the lowest values of G' and fracture stress. It was concluded that whey proteins/flavanol interactions were responsible for the poor mechanical properties of flavanol-added acidified milk gels. High-performance liquid chromatography analysis of milk sera showed that 60% of the total available monomeric flavanols was found in the serum phase from which 75% was non-associated to whey proteins. Concomitantly, >70% of flavanols with degree of polymerization >3 were found to be associated with the casein fraction.

Various types of polysaccharides are widely used in cultured dairy products. However, the interaction mechanisms, between milk proteins and these polysaccharides, are not entirely clear. To explore the interactions between uncharged and charged polysaccharides and the caseins, we used a model acid-milk-gel system, which allowed acidification to occur separately from gelation. The effect of adding uncharged dextran (DX; molecular weight ~2.0×10(6) Da) and negatively charged dextran sulfate (DS; molecular weight ~1.4×10(6) Da) to model acid milk gels was studied. Two concentrations (0.075 and 0.5%, wt/wt) of DX or DS were added to cold milk (~0°C) that had been acidified to pH values 4.4, 4.6, 4.8, or 4.9. Acidified milks containing DX or DS were then quiescently heated at the rate of 0.5°C/min to 30°C, which induced gelation, and gels were then held at 30°C for 17 h to facilitate gel development. Dynamic small-amplitude-oscillation rheology and large-deformation (shear) tests were performed. Microstructure of gels was examined by fluorescence microscopy. Gels made with a high concentration of DX gelled at a lower temperature, but after 17 h at 30°C, these gels exhibited lower storage moduli and lower yield-stress values. At pH 4.8 or 4.9 (pH values greater than the isoelectric point of caseins), addition of 0.5% DS to acidified milk resulted in lower gelation temperature. At pH 4.4 (pH values less than the isoelectric point of caseins), addition of 0.5% DS to acidified milk resulted in gels with very high stiffness values. Gels made at pH 4.8 or 4.9 with both concentrations of DS had much lower stiffness and yield-stress values than control gels. Microstructural analysis indicated that gels made at pH 4.4 with the addition of 0.5% DX exhibited large protein strands and pores, whereas gels made with 0.075% DX or the control gels had a finer protein matrix. At higher pH values (>4.4), gels made with 0.5% DX had a finer structure. At all pH values, gels made

In this study, the ability of micellar casein (MC) to interact with curcumin during acidification and to produce acid gel was investigated. Steady-state fluorescence spectroscopy of curcumin variation and fluorescence quenching of caseins upon binding with curcumin molecules were evidenced. Increasing the temperature from 20 to 35 °C enhanced MC-curcumin interactions as reflected by the increase in the binding constant from 0.6 ± 0.3 × 10(4) to 6.6 ± 0.6 × 10(4) M(-1). From changes in entropy, enthalpy and Gibbs free energy, hydrophobic interactions were proposed as major binding forces. Static fluorescence MC quenching was demonstrated for the MC-curcumin complex during acidification. From pH 7.4 to pH 5.0, the binding site numbers varied in the range from 1.25 ± 0.05 to 1.49 ± 0.05 and the binding constant kb varied from 3.9 ± 0.4 × 10(4) to 7.5 ± 0.7 × 10(4) M(-1). Small angle X-ray scattering profiles demonstrated that the MC internal structure was unchanged upon curcumin binding. The ζ-potential value of curcumin-doped MC indicated that curcumin did not modify the global charge of MC particles. Acid gelation studied by oscillation rheology and static multiple light scattering at 20 and 35 °C led to a similar behavior for native and curcumin-doped MC suspensions. For the first time, it was demonstrated that the colloidal and functional properties of MC were unchanged when doped with curcumin during acidification.

minerals to the new-born. Human alpha(s1)-casein has been reported to be much less phosphorylated than ruminant caseins, which may indicate a different function of caseins in humans. Revealing the phosphorylation pattern in human casein can thus shed light on its function. The current study found...

Full Text Available Thermoresponsive gelling materials constructed from natural and synthetic polymers can be used to provide triggered action and therefore customised products such as drug delivery and regenerative medicine types as well as for other industries. Some materials give Arrhenius-type viscosity changes based on coil to globule transitions. Others produce more counterintuitive responses to temperature change because of agglomeration induced by enthalpic or entropic drivers. Extensive covalent crosslinking superimposes complexity of response and the upper and lower critical solution temperatures can translate to critical volume temperatures for these swellable but insoluble gels. Their structure and volume response confer advantages for actuation though they lack robustness. Dynamic covalent bonding has created an intermediate category where shape moulding and self-healing variants are useful for several platforms. Developing synthesis methodology—for example, Reversible Addition Fragmentation chain Transfer (RAFT and Atomic Transfer Radical Polymerisation (ATRP—provides an almost infinite range of materials that can be used for many of these gelling systems. For those that self-assemble into micelle systems that can gel, the upper and lower critical solution temperatures (UCST and LCST are analogous to those for simpler dispersible polymers. However, the tuned hydrophobic-hydrophilic balance plus the introduction of additional pH-sensitivity and, for instance, thermochromic response, open the potential for coupled mechanisms to create complex drug targeting effects at the cellular level.

Full Text Available The major physiological function of milk is the transport of amino acids, carbohydrates, lipids and minerals to mammalian offspring. Caseins, the major milk proteins, are secreted in the form of a micelle consisting of protein and calcium-phosphate.We have analysed the role of the milk protein α-casein by inactivating the corresponding gene in mice. Absence of α-casein protein significantly curtails secretion of other milk proteins and calcium-phosphate, suggesting a role for α-casein in the establishment of casein micelles. In contrast, secretion of albumin, which is not synthesized in the mammary epithelium, into milk is not reduced. The absence of α-casein also significantly inhibits transcription of the other casein genes. α-Casein deficiency severely delays pup growth during lactation and results in a life-long body size reduction compared to control animals, but has only transient effects on physical and behavioural development of the pups. The data support a critical role for α-casein in casein micelle assembly. The results also confirm lactation as a critical window of metabolic programming and suggest milk protein concentration as a decisive factor in determining adult body weight.

The study was conducted to develop a high-performance liquid chromatographic (HPLC) method to quantify casein genetic variants (αs2-, β-, and κ-casein) in milk of homozygous individuals of Girgentana goat breed. For calibration experiments, pure genetic variants were extracted from individual milk samples of animals with known genotypes. The described HPLC approach was precise, accurate and highly suitable for quantification of goat casein genetic variants of homozygous individuals. The amount of each casein per allele was: αs2-casein A = 2.9 ± 0.8 g/L and F = 1.8 ± 0.4 g/L; β-casein C = 3.0 ± 0.8 g/L and C1 = 2.0 ± 0.7 g/L and κ-casein A = 1.6 ± 0.3 g/L and B = 1.1 ± 0.2 g/L. A good correlation was found between the quantities of αs2-casein genetic variants A and F, and β-casein C and C1 with other previously described method. The main important result was obtained for κ-casein because, till now, no data were available on quantification of single genetic variants for this protein.

The aim of this study was to investigate the role of the heat-treatment sequence of biopolymer mixtures as a formulation parameter on the acid-induced gelation of tri-polymeric systems composed of sodium caseinate (Na-caseinate), whey protein concentrate (WPC), and gum tragacanth (GT). This was studied by applying four sequences of heat treatment: (A) co-heating all three biopolymers; (B) heating the milk-protein dispersion and the GT dispersion separately; (C) heating the dispersion containing Na-caseinate and GT together and heating whey protein alone; and (D) co-heating whey protein with GT and heating Na-caseinate alone. According to small-deformation rheological measurements, the strength of the mixed-gel network decreased in the order: C>B>D>A samples. SEM micrographs show that the network of sample C is much more homogenous, coarse and dense than sample A, while the networks of samples B and D are of intermediate density. The heat-treatment sequence of the biopolymer mixtures as a formulation parameter thus offers an opportunity to control the microstructure and rheological properties of mixed gels.

Full Text Available Complex nanomicelles were made from casein and keratin through electrostatic self-assembly and transglutaminase fixation that was proved to be harmless and green. The complex nanomicelles were characterized by dynamic light scattering, scanning electron microscopy, atomic force microscopy, Fourier transform infrared spectroscopy, differential scanning calorimetry, and steady-state florescence. The results show that the complex nanomcelles acquired at the neutral pH in the mass ratio of casein to keratin 4 : 1 exhibit an anomalous sphere shape with uniform size which the diameter is about 40–70 nm. The complex nanomicelles in solution possess excellent dilution and storage stability due to the fixation and their high ζ-potential (22.8 mV. The complex nanomicelles are relatively hydrophilic and have a good potential for industrial application.

Full Text Available Abstract The genetic diversity of the casein locus in cattle was studied on the basis of haplotype analysis. Consideration of recently described genetic variants of the casein genes which to date have not been the subject of diversity studies, allowed the identification of new haplotypes. Genotyping of 30 cattle breeds from four continents revealed a geographically associated distribution of haplotypes, mainly defined by frequencies of alleles at CSN1S1 and CSN3. The genetic diversity within taurine breeds in Europe was found to decrease significantly from the south to the north and from the east to the west. Such geographic patterns of cattle genetic variation at the casein locus may be a result of the domestication process of modern cattle as well as geographically differentiated natural or artificial selection. The comparison of African Bos taurus and Bos indicus breeds allowed the identification of several Bos indicus specific haplotypes (CSN1S1*C-CSN2*A2-CSN3*AI/CSN3*H that are not found in pure taurine breeds. The occurrence of such haplotypes in southern European breeds also suggests that an introgression of indicine genes into taurine breeds could have contributed to the distribution of the genetic variation observed.

Human milk proteins provide term and preterm infants with both nutrition and protection. The objective of the present study was to examine longitudinal changes in the protein composition of term and preterm milk during the first 2 months of lactation, focusing on protein phosphorylation and glycosylation. Using gel electrophoresis, the relative concentration and glycosylation status of lactoferrin, secretory Ig A, β-casein, α-lactalbumin, serum albumin, bile salt-stimulated lipase, xanthine oxidoreductase, tenascin and macrophage mannose receptor 1 were measured in milk collected on days 7, 10, 14, 18, 21, 28 and 60 postpartum from preterm mothers (28-32 weeks gestation, n 17). The phosphorylation status of β-casein was also investigated. To determine if these variables differ in term and preterm milk, samples from term mothers (38-41 weeks gestation, n 8) collected on days 7, 14 and 30 of lactation were also analysed. The concentration of the abundant milk proteins decreased during lactation in term and preterm milk (P protein glycosylation was observed, except for the glycoproteins serum albumin and tenascin. The phosphorylation of β-casein varied significantly between term and preterm milk. Further investigation is required to determine whether these modifications affect protein function and are clinically important to preterm infants.

Background It has been suggested that peptides from gluten and casein may have a role in the origins of autism and that the physiology and psychology of autism might be explained by excessive opioid activity linked to these peptides. Research has reported abnormal levels of peptides in the urine and cerebrospinal fluid of people with autism. Objectives To determine the efficacy of gluten and/or casein free diets as an intervention to improve behaviour, cognitive and social functioning in individuals with autism. Search methods The following electronic databases were searched: CENTRAL(The Cochrane Library Issue 2, 2007), MEDLINE (1966 to April 2007), PsycINFO (1971 to April 2007), EMBASE (1974 to April 2007), CINAHL (1982 to April 2007), ERIC (1965 to 2007), LILACS (1982 to April 2007), and the National Research register 2007 (Issue1). Review bibliographies were also examined to identify potential trials. Selection criteria All randomised controlled trials (RCT) involving programmes which eliminated gluten, casein or both gluten and casein from the diets of individuals diagnosed with an autistic spectrum disorder. Data collection and analysis Abstracts of studies identified in searches of electronic databases were assessed to determine inclusion by two independent authors The included trials did not share common outcome measures and therefore no meta-analysis was possible. Data are presented in narrative form. Main results Two small RCTs were identified (n = 35). No meta-analysis was possible. There were only three significant treatment effects in favour of the diet intervention: overall autistic traits, mean difference (MD) = −5.60 (95% CI −9.02 to −2.18), z = 3.21, p=0.001 (Knivsberg 2002) ; social isolation, MD = −3.20 (95% CI −5.20 to 1.20), z = 3.14, p = 0.002) and overall ability to communicate and interact, MD = 1.70 (95% CI 0.50 to 2.90), z = 2.77, p = 0.006) (Knivsberg 2003). In addition three outcomes showed no significant difference between the

The current research reports partial characterization of the caseins and α-lactalbumin (α-LA) of the African elephant with proposed unique structure-function properties. Extensive research has been carried out to understand the structure of the casein micelles. Crystallographic structure elucidation of caseins and casein micelles is not possible. Consequently, several models have been developed in an effort to describe the casein micelle, specifically of cow milk. Here we report the characterization of African elephant milk caseins. The κ-caseins and β-caseins were investigated, and their relative ratio was found to be approximately 1:8.5, whereas α-caseins were not detected. The gene sequence of β-casein in the NCBI database was revisited, and a different sequence in the N-terminal region is proposed. Amino acid sequence alignment and hydropathy plots showed that the κ-casein of African elephant milk is similar to that of other mammals, whereas the β-casein is similar to the human protein, and displayed a section of unique AA composition and additional hydrophilic regions compared with bovine caseins. Elephant milk is destabilized by 62% alcohol, and it is speculated that the β-casein characteristics may allow maintenance of the colloidal nature of the casein micelle, a role that was previously only associated with κ-casein. The oligosaccharide content of milk was reported to be low in dairy animals but high in some other species such as humans and elephants. In the milk of the African elephant, lactose and oligosaccharides both occur at high levels. These levels are typically related to the content of α-LA in the mammary gland and thus point to a specialized carbohydrate synthesis, where the whey protein α-LA plays a role. We report the characterization of African elephant α-LA. Homology modeling of the α-LA showed that it is structurally similar to crystal structures of other mammalian species, which in turn may be an indication that its functional

The time-dependent evolution of the phase-separated microstructure of a caseinate-stabilised emulsion containing xanthan gum added before emulsification has been investigated by confocal laser scanning microscopy, image analysis and rheology. Moderately low levels of xanthan addition lead to depletion flocculation and gravity-induced phase separation. Increasing the polysaccharide concentration causes immobilisation of the microstructure due to an increase in the local viscoelasticity: that is, the emulsion structure cannot easily rearrange to expel xanthan-enriched aqueous serum phase because a weak gel-like network is generated. The effect of xanthan on the evolving microstructure of phase-separated regions, which reflects indirectly the local emulsion micro-rheology, has been estimated from image analysis of time sequences of confocal micrographs. A comparison has been made between object shape analysis using four different shape descriptors. The roundness parameter has been found to be a convenient descriptor for reliably quantifying the structural change in terms of the relaxation rate of xanthan-rich aqueous drops. The Taylor parameter has been used to link the kinetics of drop relaxation to the time-dependent small-deformation rheological behaviour. The analysis of the combined experimental data reveals the difficulty of relating the evolving microstructure to bulk rheological measurements.

Full Text Available Introduction: Casein is a milk protein that has the same number of positive and negative charges in a pH around 4.7. This characteristic is called the isoelectric point (pI. The pI varies from protein to protein and it depends on the charges of the lateral chain of the constituent aminoacids. At this pH value the protein has its minimum solubility since the net charge is zero and the repulsion between molecules is decreased. Furthermore, the electrostatic interaction occurs between the protein molecules. Thus, they form clumps which tend to precipitate. On the other hand, when they are placed in a solution whose pH is above or below of its pI, the protein molecules have respectively a negative or positive net charge, with strong repulsion between themselves and great interaction with the solvent (water. Objective: This learning object presents a simulation of a laboratory practice for the determination of casein isoelectric point. Materials and Methods: Cartoons were planned in order to show the methodology procedures and biochemical fundamentals. Animations were developed with the aid of the Adobe ® Flash 8 software associated with logic programming. Results: The simulation consists of six steps that reproduce the activities performed in the laboratory. Among them, the user can observe different degrees of turbidity and / or precipitation of casein in solutions with different pH values, he can measure these values with paper indicator strips and then determine the pI of this protein. Conclusions: This learning object was tested by students of Biochemistry I, Pharmacy course since 2012/2. The navigation features, design, interaction, interactivity were considered excellent by 80% of students indicating that this object can be used as an interesting tool to assist the teaching and learning of basic biochemistry. Available at:www.ufrgs.br/gcoeb/PontoIsoleletricoDaCaseina/PontoIsoleletricoDaCaseina.swf

By using seven endonucleases and four bovine cDNA probes specific for alpha S1-, alpha S2-, beta-, and kappa-casein genes, nine restriction fragment length polymorphisms (RFLPs) have been found in the sheep orthologous DNA regions. In contrast to the low level of variation observed at the protein level, these DNA polymorphisms determine a high level of heterozygosity and, therefore, represent useful tools for genetic analyses since they can also be obtained without the need for gene expression. In fact, informative matings suggest that in sheep, as in cattle, the four loci are linked.

Introduction: Casein is a milk protein that has the same number of positive and negative charges in a pH around 4.7. This characteristic is called the isoelectric point (pI). The pI varies from protein to protein and it depends on the charges of the lateral chain of the constituent aminoacids. At this pH value the protein has its minimum solubility since the net charge is zero and the repulsion between molecules is decreased. Furthermore, the electrostatic interaction occurs between the prote...

Previous studies show increased antibody levels to bovine casein in some individuals with schizophrenia. The immunogenicity of specific domains of bovine casein varies among people with milk sensitivities and thus could vary among different neuropsychiatric disorders. Using ELISAs and immunoblotting

insulin, and insulin secretion estimated as the plasma C-peptide concentration in overweight adolescents. Overweight adolescents (n = 203) aged 12–15 y with a BMI of 25.4 ± 2.3 kg/m2 (mean ± SD) were randomized to 1 L/d of skim milk, whey, casein, or water for 12 wk. All milk drinks contained 35 g protein...... and with the water and pretest control groups. The plasma C-peptide concentration increased from baseline to wk 12 in the whey and casein groups and increments were greater than in the pretest control (P ... that high intakes of skim milk, whey, and casein increase BAZs in overweight adolescents and that whey and casein increase insulin secretion. Whether the effect on body weight is primary or secondary to the increased insulin secretion remains to be elucidated....

The effect of caseinate and soy protein in the diet on the mutagenicity induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) was assessed in-vivo and ex-vivo in the DNA-repair host-mediated assay and liquid suspension assay, respectively. Of the two proteins only casein showed a strong antimutagen

The physical properties of ice cream are mainly determined by the processing and the ingredients. Milk (powder) is one of the ingredients and ice cream thus contains casein, the major milk protein. A large proportion of casein in ice cream is present in the plasma phase of ice cream. Since the behav

Whey protein and casein were hydrolyzed with 11 commercially available enzymes. Foam properties of 44 samples were measured and were related to biochemical properties of the hydrolysates using statistical data analysis. All casein hydrolysates formed high initial foam levels, whereas whey hydrolysat

Polymorphism of casein genes was studied in half-sib families of artificial insemination bulls of the Finnish Ayrshire dairy breed. Ten grandsires and 300 of their sons were genotyped for the following polymorphisms: alpha s1-casein (B, C), beta-casein (A1, A2), the microsatellite within the kappa-casein gene (ms5, ms4) and kappa-casein (A, B, E). Nine different combinations of these alleles, casein haplotypes, were found. Associations between casein haplotypes and milk production traits (milk and protein yield, fat and protein percentage and milking speed) were studied with ordinary least-squares analysis to find a direct effect of the haplotypes or an association within individual grandsire families using the granddaughter design. Estimated breeding values of sons were obtained from cow evaluation by animal model. No direct effect of the casein haplotypes on the traits was found. Within grandsire families, in one out of four families the chromosomal segment characterized by haplotype 3 (B-A2-ms4-A) was associated with an increase in milk yield (P < 0.01) and a decrease in fat percentage (P < 0.01) when contrasted with haplotype 8 (B-A1-ms4-E). The results provide evidence that in the Finnish Ayrshire breed at least one quantitative trait locus affecting the genetic variation in yields traits is segregating linked to either haplotype 3 (B-A2-ms4-A) or 8 (B-A1-ms4-E).

Growth assays were performed to determine lysine bioavailability for kittens and rats in untreated and heated casein; these values were compared with estimates obtained with an in vitro method. Body weight, food intake, nitrogen and dry matter digestibility, and plasma lysine were determined during an 80-day growth trial using kittens (n = 16). Body weight and food intake were determined during a 21-day growth trial using weanling rats (n = 80). The growth data showed bioavailable lysine to be 102.4% and 100.2% (for untreated casein) and 66.1% and 51.7% (for heated casein) for kittens and rats, respectively. There was no relationship between plasma lysine and dietary lysine concentrations for kittens. There were no significant differences in nitrogen or dry matter digestibility among diets for kittens. The chemically reactive lysine content of untreated casein was 99.6%, and of heated casein was 67.1%. Heat treatment of casein resulted in significantly decreased lysine bioavailability as estimated by all methods. For untreated casein, both growth assays showed good agreement with the in vitro method for available lysine. For heated casein, the rat growth assay significantly underestimated bioavailable lysine as determined in kittens while the in vitro method closely approximated this value for the cat.

Bovine chymosin is an aspartic protease that selectively cleaves the milk protein κ-casein. The enzyme is widely used to promote milk clotting in cheese manufacturing. We have developed models of residues 97-112 of bovine κ-casein complexed with bovine chymosin, using ligand docking, conformational...

Full Text Available CASEIN, a protein found in milk of several species, is divided into different chains from 19 to 25 kDa. Casein is also considered as a source of amino acids and generating peptides with biological activities such as opiate, immunostimulating, antibacterial, peptidase inhibitors, among others.

The agreed biological function of the casein micelles in milk is to carry minerals (calcium, magnesium, and phosphorus) from mother to young along with amino acids for growth and development. Recently, native and modified casein micelles were used as encapsulating and delivery agents for various hydrophobic low-molecular-weight probes. The ability of modified casein micelles to bind certain probes may derive from the binding affinity of native casein micelles. Hence, a study with milk from single cows was conducted to further elucidate the association of hydrophobic molecules into native casein micelles and further understand their biological function. Hydrophobic and hydrophilic extraction followed by ultraperformance liquid chromatography-high resolution mass spectrometry analysis were performed over protein fractions obtained from size exclusion fractionation of raw skim milk. Hydrophobic compounds, including phosphatidylcholine, lyso-phosphatidylcholine, phosphatidylethanolamine, and sphingomyelin, showed strong association exclusively to casein micelles as compared with whey proteins, whereas hydrophilic compounds did not display any preference for their association among milk proteins. Further analysis using liquid chromatography-tandem mass spectrometry detected 42 compounds associated solely with the casein-micelles fraction. Mass fragments in tandem mass spectrometry identified 4 of these compounds as phosphatidylcholine with fatty acid composition of 16:0/18:1, 14:0/16:0, 16:0/16:0, and 18:1/18:0. These results support that transporting low-molecular-weight hydrophobic molecules is also a biological function of the casein micelles in milk.

Interfacial rheological properties and their suitability for foam production and stability of two vegetable proteins were studied and compared to ß-casein. Proteins used ranged from flexible to rigid/globular in the order of ß-casein, gliadin and soy glycinin. Experiments were performed at pH 6.7. N

Full Text Available Aim: Sheep milk-born bioactive peptides have been found to exhibit various physiological activities. The present work was designed with the aim to evaluate the effect of boiling on antidiabetic property of peptides derived from sheep milk caseinate on hydrolysis with three different proteases. Materials and Methods: In this investigation, casein prepared from raw and boiled sheep milk was hydrolyzed by three commercially available proteases (trypsin, pepsin, and chymotrypsin. These hydrolysates collected at different hydrolysis times (60, 120, 160, and 240 min were assayed for their antidiabetic activity. Results: Among the three different enzyme hydrolysates, casein treated with chymotrypsin shows the highest antidiabetic activity among other enzymes. While the antidiabetic effect of raw milk-derived casein hydrolysates always exceeds than effect shown by boiled milk casein hydrolysates. Conclusion: The result obtained hence shows that the effect of boiling on the properties of bioactive peptides released during different enzyme digestion depends largely on the enzymatic formulation used and treatment conditions. Chymotrypsin treatment of raw casein yields peptides with maximum antidiabetic activity as compared to pepsin and trypsin. Moreover, the peptides produced after enzymatic treatment of boiled casein show reduced antidiabetic properties. Therefore, enzymatically treated raw milk casein hydrolysates may be used as effective nutritional supplements for diabetic patients, as it causes a significant inhibition of α-amylase activity.

Interfacial rheological properties and their suitability for foam production and stability of two vegetable proteins were studied and compared to β-casein. Proteins used ranged from flexible to rigid/globular in the order of β-casein, gliadin and soy glycinin. Experiments were performed at pH 6.7. N

Casein micelles contribute to the physicochemical properties of milk and may also influence its functionality. At present, however, there is an incomplete understanding of the casein micelle associated proteins and its diversity among the milk obtained from different species. Therefore, milk samples were collected from seven dairy animals groups, casein fractions were prepared by ultracentrifugation and their constituent proteins were identified by liquid chromatography tandem mass spectrometry. A total of 193 distinct proteins were identified among all the casein micelle preparations. Protein interaction analysis indicated that caseins could interact with major whey proteins, including β-lactoglobulin, α-lactalbumin, lactoferrin, and serum albumin, and then whey proteins interacted with other proteins. Pathway analysis found that the peroxisome proliferator-activated receptor signaling pathway is shared among the studied animals. Additionally, galactose metabolism pathway is also found to be commonly involved for proteins derived from camel and horse milk. According to the similarity of casein micelle proteomes, two major sample clusters were classified into ruminant animals (Holstein and Jersey cows, buffaloes, yaks, and goats) and non-ruminants (camels and horses). Our results provide new insights into the protein profile associated with casein micelles and the functionality of the casein micelle from the studied animals. PMID:28240229

The physical properties of ice cream are mainly determined by the processing and the ingredients. Milk (powder) is one of the ingredients and ice cream thus contains casein, the major milk protein. A large proportion of casein in ice cream is present in the plasma phase of ice cream. Since

Whey protein and casein were hydrolyzed with 11 commercially available enzymes. Foam properties of 44 samples were measured and were related to biochemical properties of the hydrolysates using statistical data analysis. All casein hydrolysates formed high initial foam levels, whereas whey hydrolysates differed in their foam-forming abilities. Regression analysis using the molecular weight distribution of whey hydrolysates as predictors showed that the hydrolysate fraction containing peptides of 3-5 kDa was most strongly related to foam formation. Foam stability of whey hydrolysates and of most casein hydrolysates was inferior to that of the intact proteins. The foam stability of casein hydrolysate foams was correlated to the molecular weight distribution of the hydrolysates; a high proportion of peptides >7 kDa, composed of both intact casein and high molecular weight peptides, was positively related to foam stability.

The effects of temperature,ionic strength,and enzymatic hydrolysis on the average hydrodynamic radius (Rh) of casein micelles in phosphate buffer were studied by using dynamic light scattering.The results showed that the average Rh value of casein mieelles decreased irreversibly during the heating,decreased with the increase of ionic strength in lower ionic strength solution (less than 0.05 tool/L),but opposite in higher ionic strength solution (above 0.1 tool/L).The Rh value of casein increased rapidly during the process of enzymatic hydrolysis,and the structural model of casein micelles in the enzymatic hydrolysis process was also proposed,i.e.the casein micelle changed from compact sphere into unfolded and regularly flocky peptides.

The reaction between p-benzoquinone (PBQ) and casein (protein) in the solid state was studied. After reaction with protein, the FT-IR spectra showed a new band at 1215 cm(-1), with an intense sign attributed for casein/PBQ product. An optimum amount of PBQ for determination of the product when mixed with casein was 100mg and a reaction time of 30 min. The product was stable for a period of 24h after the reacting by heating. The interaction of PBQ with casein was investigated by FT-IR, reflectance, Raman and EPR spectroscopies. The reaction between PBQ and casein in the solid state yields a radical species (p-benzosemiquinone) that it is stabilized by the presence of p-biphenolate and p-benzoquinone species in the solid state. Copyright (c) 2009 Elsevier B.V. All rights reserved.

A new biodegradable flocculant was prepared from a common and inexpensive protein. Milk casein was methylated in a 0.05 M HCl methyl alcohol solution at room temperature. The methylated milk casein (MeCS), having a methylation degree of 81%, was applied to the separation or flocculation of diatomite in seawater (pH 8.1+/-0.1) at room temperature (18-23 degrees C). The flocculating ability of MeCS was evaluated by a sedimentation balance method (cumulative measurement method). The diatomite suspension was effectively flocculated by the addition of a small amount of MeCS (0.25 wt% of the diatomite weight). The results of sedimentation analysis showed that the size-frequency curve had a very sharp and high peak; thus the diatomite floc formed by MeCS had a rather uniform size. The settling velocity of diatomite floc at the appropriate MeCS dosages (0.25-2 wt% of diatomite) was about 3 x 10(-3) ms(-1).

@@ 1Introduction In present trade mark adhesives for beer bottle labeling are mostly prepared based on starch[1] ,polyvinyl formal[2] and their derivants[3-5] ,but they can only be applied to the labeling speed of 20 000 bottle/h and their water proofness are not satisfactory, especially when the water temperature is 0℃ below. The application of metal foil requires that sealing and labeling must be finished at the same time, but different penetrating speed of water in metal foil and paper label causes different drying speed, and also, the labeling speed and need of iced beer are increasing now, so these changes result in more strict requirements for adhesive.In this paper the chelating casein adhesive was obtained with casein as main moldings, urea as dispersing agent, starch as auxiliary, and ZnSO4 as cross-linked agent. The effects of dispersing agent, cross-linked agent and starch on the performances of adhesive were investigated.

Ultrastructure of lactating bovine and rat mammary epithelial cells was studied with emphasis on secretory vesicle interactions. In the apical zone of the cell, adjacent secretory vesicles formed ball and socket configurations at their points of apposition. Similar configurations were formed between plasma membrane and secretory vesicle membrane. These structures may be formed by the diffusion of water between vesicles with different osmotic potentials. Frequently, vesicular chains consisting of 10 or more linked secretory vesicles were observed. Prior to the exocytotic release of casein micelles, adjacent vesicles fused through fragmentation of the ball and socket membrane. These membrane fragments and the casein micelles appeared to be secreted into the alveolar lumen after passing from one vesicle into another and finally through a pore in the apical plasma membrane. Emptied vesicular chains appeared to collapse and fragmentation of their membrane was observed. Based on these observations, we suggest that most vesicular membrane does not directly contact or become incorporated into the plasma membrane during secretion of the nonfat phase of milk.

Purpose Despite most epidemiologic studies reporting that an increase in milk intake affects the growth of prostate cancer, the results of experimental studies are not consistent. In this study, we investigated the proliferation of prostate cancer cells treated with casein, the main protein in milk. Materials and Methods Prostate cancer cells (LNCaP and PC3), lung cancer cells (A459), stomach cancer cells (SNU484), breast cancer cells (MCF7), immortalized human embryonic kidney cells (HEK293), and immortalized normal prostate cells (RWPE1) were treated with either 0.1 or 1 mg/mL of α-casein and total casein extracted from bovine milk. Treatments were carried out in serum-free media for 72 hours. The proliferation of each cell line was evaluated by an 3-(4,5-Dimethyl-thiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Results α-Casein and total casein did not affect the proliferations of RWPE1, HEK293, A459, SNU484, MCF7, HEK293, or RWPE1 cells. However, PC3 cells treated with 1 mg/mL of α-casein and casein showed increased proliferation (228% and 166%, respectively), and the proliferation of LNCaP cells was also enhanced by 134% and 142%, respectively. The proliferation mechanism of α-casein in PC3 and LNCaP cells did not appear to be related to the induction of Insulin-like growth factor-1 (IGF-1), since the level of IGF-1 did not change upon the supplementation of casein. Conclusions The milk protein, casein, promotes the proliferation of prostate cancer cells such as PC3 and LNCaP. PMID:25237656

Testosterone nasal gel is used to treat symptoms of low testosterone in men who have hypogonadism (a condition in which the ... does not produce enough natural testosterone). Testosterone nasal gel is used only for men with low testosterone ...

The aim of this study is to determine the effects of plasticizer hydrogen bonding capability and chain length on the molecular structure of sodium caseinate (NaCAS), in NaCAS/glycerol and NaCAS/polyethylene glycol 400 (PEG) systems. Both solution and film phases were investigated. Glycerol and PEG reduced the viscosity of aqueous NaCAS, with the latter having a greater effect. This was explained in terms of protein/plasticizer aggregate size and changes to the conformation of the caseinate chain. In the film phase, glycerol caused more pronounced changes to the film tensile strength compared with PEG. However, the effect of glycerol on film water vapor permeability was smaller. These observations are attributed to the differences in plasticizer size and hydrogen bonding strength that controls the protein-plasticizer and protein-protein interactions in the films. Glass transition calculations from the tensile strength data indicate that the distribution of bonding interactions is more homogeneous in NaCAS/PEG films than in NaCAS/glycerol films.

Numerous peptides generated by triptic hydrolysis of micellar casein and β-casein from bovine milk have been reported to exhibit bio-functional or techno-functional activities. In this study, the focus was placed on the quantification of eleven potential functional peptides generated by enzymatic hydrolysis with trypsin. The identity of the target peptides was verified by LC-ESI-MS/MS and a reverse phase high performance liquid chromatography method (RP-HPLC) was established to quantify the peptides. The quantification was based on calibration curves drawn using synthesised purified peptides. The regression analysis comparing >30 standard measurements of each peptide standard showed a regression coefficient R(2)>0.9958. The inter-day repeatability of the quantification method was always within a relative error of 0.2-6.5%, while the relative error of the accuracy for reproducibility was in the range of 0.1-2.4%. The established method was successfully applied for the quantitative analysis of the eleven functional peptides in different dairy fractions generated by cross-flow ultrafiltration.

The rennet-induced clotting of milk was studied under various conditions. The kinetics of the enzymic and aggregation reactions was analysed separately and, where possible, related to the physico-chemical properties of the casein micelle and its environment.

The interaction of bisdemethoxycurcumin (BDMC), as one of the main active component of turmeric (Curcuma longa L.), with bovine β-casein nanoparticle, as an efficient drug carrier system, was investigated using steady-state fluorescence spectroscopy and molecular docking calculations. Results of fluorescence quenching experiments, Forster energy transfer measurements and molecular docking calculations suggested that BDMC bind to the hydrophobic core of β-casein via formation of 3 hydrogen bonds and several vander Waals contacts that represented the encapsulation of BDMC in β-casein micelle nanoparticles. The binding parameters including number of substantive binding sites and the binding constants were evaluated by fluorescence quenching method. Additionally, the cytotoxicity of free BDMC and BDMC-β-casein complex in human breast cancer cell line MCF7 was evaluated in vitro. The study revealed the higher cytotoxic effects of encapsulated BDMC on MCF7 cells compared to equal dose of free BDMC. -- Highlights: • BDMC binds to the hydrophobic core of β-casein. • The effective encapsulation of BDMC in β-casein micelle nanoparticles was shown. • Enhanced cytotoxicity was observed for encapsulated BDMC in β-casein nanoparticles.

Whey protein concentrate (WPC 80) and sodium caseinate were hydrolyzed by Protamex to 5%, 10%, 15%, and 20% degree of hydrolysis (DH). WPC 80, sodium caseinate and their hydrolysates were then analyzed, compared and evaluated for their nutritional qualities. Their chemical composition, protein solubility, amino acid composition, essential amino acid index (EAA index), biological value (BV), nutritional index (NI), chemical score, enzymic protein efficiency ratio (E-PER) and in vitro protein digestibility (IVPD) were determined. The results indicated that the enzymatic hydrolysis of WPC 80 and sodium caseinate by Protamex improved the solubility and IVPD of their hydrolysates. WPC 80, sodium caseinate and their hydrolysates were high-quality proteins and had a surplus of essential amino acids compared with the FAO/WHO/UNU (1985) reference standard. The nutritive value of WPC 80 and its hydrolysates was superior to that of sodium caseinate and its hydrolysates as indicated by some nutritional parameters such as the amino acid composition, chemical score, EAA index and predicted BV. However, the E-PER was lower for the WPC hydrolysates as compared to unhydrolyzed WPC 80 but sodium caseinate and its hydrolysates did not differ significantly. The nutritional qualities of WPC 80, sodium caseinate and their hydrolysates were good and make them appropriate for food formulations or as nutritional supplements.

Human milk contains many bioactive proteins that are likely to support the early development of the newborn. The aim of this study was to identify whether there are specific minor proteins associated with the human milk casein micelle prepared by the acid precipitation method. Protein identification was performed by liquid chromatography tandem mass spectrometry analysis. Eighty-two proteins were identified in the casein micelle, 18 of which are not present in their whey compartment. Thirty-two of these proteins specifically associated with the casein micelle have not previously been identified in human milk or colostrum. Proteins involved in immune function comprised the major part (28%) of total proteins, and another significant part is involved in metabolism/energy production (22%). Most of the proteins were of extracellular or cytoplasmic origin (accounting for 50 and 29%, respectively). This study indicates that various soluble proteins should be considered as part of the casein compartment, prepared by the acid precipitation method. The data provide new insight not only into the proteomic profile of the human milk casein micelle and its physiological significance, but also into the proper proportion of casein and casein-associated proteins to use in infant formula.

Whey protein concentrate (WPC 80) and sodium caseinate were hydrolyzed by Protamex to 5%, 10%, 15%, and 20% degree of hydrolysis (DH). WPC 80, sodium caseinate and their hydrolysates were then analyzed, compared and evaluated for their nutritional qualities. Their chemical composition, protein solubility, amino acid composition, essential amino acid index (EAA index), biological value (BV), nutritional index (NI), chemical score, enzymic protein efficiency ratio (E-PER) and in vitro protein digestibility (IVPD) were determined. The results indicated that the enzymatic hydrolysis of WPC 80 and sodium caseinate by Protamex improved the solubility and IVPD of their hydrolysates. WPC 80, sodium caseinate and their hydrolysates were high-quality proteins and had a surplus of essential amino acids compared with the FAO/WHO/UNU (1985) reference standard.The nutritive value of WPC 80 and its hydrolysates was superior to that of sodium caseinate and its hydrolysates as indicated by some nutritional parameters such as the amino acid composition, chemical score, EAA index and predicted BV. However, the E-PER was lower for the WPC hydrolysates as compared to unhydrolyzed WPC 80 but sodium caseinate and its hydrolysates did not differ significantly. The nutritional qualities of WPC 80, sodium caseinate and their hydrolysates were good and make them appropriate for food formulations or as nutritional supplements.

Milk protein of farm animals is difficult to isolate because of the presence of casein micelles, which are hard to separate from whey by using centrifugation or filtration. Insoluble casein micelles also create an obstacle for purification instruments to operate efficiently. The conventional method, to precipitate caseins by lowering pH to 4.6 and then recover the whey fraction for further purification using chromatography techniques, is not applicable to proteins having an isoelectric point similar to caseins. In addition, the acid condition used for casein removal usually leads to significantly poor yields and reduced biological activities. In this study, a novel method of precipitating caseins under neutral or weak acidic conditions is presented. The method employs a phosphate salt and a freeze-thaw procedure to obtain a casein-free whey protein fraction. This fraction contains more than 90% yield with little loss of bioactivity of the target protein, and is readily available for further chromatographic purification. This method was successfully applied to purify recombinant human factor IX and recombinant hirudin from the milk of transgenic pigs in the presented study. It is an efficient pretreatment approach prior to chromatographic purification of milk protein from farm animals and particularly of great value to collect those recombinants secreted from transgenic livestock.

The association of triclosan (TCS), a widely used hydrophobic compound, to the bovine casein micelle is investigated in this study. The use of high-pressure homogenization (HPH) at 0, 100, 200, and 300 MPa was introduced as a method for the dissociation of casein micelles in a skim milk/ethanol solution (1: 1, v/v) in the presence of TCS at 20, 80, and 160 mg/L where ethanol evaporation served as the final step for TCS association to caseins. The majority of TCS (over 80%) was associated with the caseins regardless of initial TCS concentration or applied pressure. TCS association to caseins was enhanced by 30% with continued pressurization to 300 MPa. Micellar dissociation and reassociation was found to be an irreversible process as evidenced by microscopy images. Pressurization to 300 MPa resulted in the formation of an integrated protein network of casein proteins and noncovalently linked whey proteins where the solubility of TCS was enhanced up to 40 times its reported water solubility at the highest initial TCS level of 160 mg/L. Reformed micelles exhibited Newtonian flow behavior at all pressure levels. This study provides evidence for the solubility enhancing quality of TCS through the solvent-mediated pressure/shear-induced dissociation of casein proteins.

The aim of this study was to identify the molecular genetic origin underlying the I variant of alpha(s1)-casein and to develop a DNA-based test for this polymorphism as a tool for genetic analyses independent of milk sample testing. All coding exons and flanking regions of the alpha(s1)-casein gene were sequenced in DNA samples from cattle of known alpha(s1)-casein genotypes (BI, CI, II, CC), determined by isoelectric focusing of milk samples. A nucleotide substitution (A>T) in exon 11 (g.19836A>T) leads to the exchange of Glu with Asp at amino acid position 84 of the mature protein (p.Glu84Asp) and perfectly co-segregated with the presence of the alpha(s1)-casein I variant in the milk of the analysed animals. Genotyping of a total of 680 DNA samples from 31 Bos taurus and Bos indicus cattle breeds and from Bos grunniens, Bison bison and Bison bonasus by restriction fragment length polymorphism analysis revealed the occurrence of Asp at position 84 at low frequencies in Bos taurus and Bos indicus breeds and established its origin from the alpha(s1)-casein C variant (p.Glu192Gly). Ten different intragenic haplotypes in the gene region from intron 8 to intron 12 were observed by sequencing, of which two occurred in Bison bison and one in Bison bonasus only. Using available casein gene complex information, an association of Asp at position 84 to beta-casein A(2) and kappa-casein B was shown in the Bos indicus breed Banyo Gudali. Taken together, we can postulate that the alpha(s1)-casein variant I is caused by a non-synonymous nucleotide substitution in exon 11 of the gene and that it originated within Bos indicus and spread to Bos taurus subsequently.

Milk is a complex mixture of various components which include proteins, carbohydrates, lipids and minerals. Milk proteins can be divided into two classes: whey proteins (20%) and caseins (80% of total milk protein). As thermal treatment is a traditional method of food preservation, interactions...... which can improve stability and safety of food. Traditional methods of food preservation and sterilization require high temperature, which may result in undesirable changes such as change of color or texture. Development of novel food processing methods, which allow to create new milk-based products...... with novel textures, is highly valuable to the dairy industry. Understanding interactions that undergo during food processing is necessary for proper design of process. For example control of aggregation is required to achieve desired texture of the final product....

Full Text Available Gel becomes an important class of soft materials since it can be seen in a wide variety of the chemical and the biological systems. The unique properties of gel arise from the structure, namely, the three-dimensional polymer network that is swollen by a huge amount of solvent. Despite the small volume fraction of the polymer network, which is usually only a few percent or less, gel shows the typical properties that belong to solids such as the elasticity. Gel is, therefore, regarded as a dilute solid because its elasticity is much smaller than that of typical solids. Because of the diluted structure, small molecules can pass along the open space of the polymer network. In addition to the viscous resistance of gel fluid, however, the substance experiences resistance due to the polymer network of gel during the transport process. It is, therefore, of importance to study the diffusion of the small molecules in gel as well as the flow of gel fluid itself through the polymer network of gel. It may be natural to assume that the effects of the resistance due to the polymer network of gel depends strongly on the network structure. Therefore, detailed study on the transport processes in and through gel may open a new insight into the relationship between the structure and the transport properties of gel. The two typical transport processes in and through gel, that is, the diffusion of small molecules due to the thermal fluctuations and the flow of gel fluid that is caused by the mechanical pressure gradient will be reviewed.

Full Text Available In bovine species, β-casein (β-CN is characterized by genetic polymorphism. The two most common protein variants are β-CN A2 (the original one and A1, differing from A2 for one amino acid substitution (Pro67 to His67. Several bioactive peptides affecting milk nutritional properties can originate from β-CN. Among them, β-casomorphin-7 (BCM7 ranging from amino acid 60 to 66 can be released more easily from β-CN variants carrying His67 (A1 type instead of Pro67 (A2 type. Nowadays, “A2 milk” is produced in different countries claiming its potential benefits in human health. The aim of this study was to further develop and apply an isoelectric focusing electrophoresis (IEF method to bulk and individual milk samples in order to improve its use for β-CN studies. We succeeded in identifying A2 milk samples correctly and quantifying the percentage of A2, A1, and B variants in bulk samples not derived from A2 milk as well as in individual milk samples. The method allows us to quantify the relative proportion of β-CN variants in whole milk without eliminating whey protein by acid or enzymatic precipitation of caseins. The aim of this study was also to study the different behavior of β-CN and β-lactoglobulin (β-LG in the presence of trichloroacetic acid (TCA. The higher sensitivity of β-CN to TCA allows quantifying β-CN variants after TCA fixation because β-LG is not visible. Monitoring β-CN variation in cattle breeds is important in order to maintain a certain balance between Pro67 and His67 in dairy products. Overall, the debate between A1 and A2 milk needs further investigation.

Restriction fragment length polymorphism (RFLP) of ovine casein genes was investigated. Genomic DNA from 56 rams was digested with 10 restriction endonucleases and Southern blots probed with the four ovine casein cDNAs (alpha s1-, beta-, alpha s2- and kappa-Cn). Five enzymes, namely, BglI, PvuII, RsaI, TaqI and HindIII revealed nine different RFLPs. The inheritance of six of these polymorphisms was studied by segregation analysis of gametes in nine rams' families, and each of them could be related to the existence of alleles at the relevant casein locus. A close linkage between the four ovine casein genes was demonstrated since no recombination within the four pairs of loci examined, alpha s1-beta-Cn, alpha s1-kappa-Cn, beta-kappa-Cn and alpha s2-kappa-Cn, was observed in the progeny of double heterozygous rams. The casein genes are thus clustered in the ovine species as in the case of other mammals.

The most polymorphic milk protein gene is β-casein; 13 protein variants are known in cattle. Milk protein genetic polymorphism has received considerable research interest in recent years because of possible associations between milk protein and economically important traits in livestock. The present study was undertaken to explore the genetic polymorphisms in exon 7 of β-casein and exon 4 of κ-casein genes in Arunachali yaks (Bos grunniens), Sahiwal (Bos indicus) cattle, malpura sheep (Ovis aries) and Gaddi goat (Capra hircus). Results of the study revealed presence of 11 SNP variants in all livestock species. Four SNPs were observed in Bos indicus; two SNPs in Bos grunniens; three SNPs in Ovis aries and three SNPs in Capra hircus. These variations are found to be synonymous in nature as these variations do not result in their corresponding amino acids. A total of five polymorphic sites have been described at the κ-casein (CSN3) locus in the Indian domestic Gaddi goat (Capra hircus) when compared with exotic goat (X60763) while sequence analysis of κ-casein gene in sheep showed three novel nucleotide changes in malpura sheep when compared with the exotic sheep (AY237637). These results highlight the importance of taking into consideration the CSN3 SNPs when performing selection for milk composition in dairy livestock breeds.

Multicomponent homogeneous, ultrapure noncrystalline gels/gel derived glasses are promising batch materials for the containerless glass melting experiments in microgravity. Hence, ultrapure, homogeneous gel precursors could be used to: (1) investigate the effect of the container induced nucleation on the glass forming ability of marginally glass forming compositions; and (2) investigate the influence of gravity on the phase separation and coarsening behavior of gel derived glasses in the liquid-liquid immiscibility zone of the nonsilicate systems having a high density phase. The structure and crystallization behavior of gels in the SiO2-GeO2 as a function of gel chemistry and thermal treatment were investigated. As are the chemical principles involved in the distribution of a second network former in silica gel matrix being investigated. The procedures for synthesizing noncrystalline gels/gel-monoliths in the SiO2-GeO2, GeO2-PbO systems were developed. Preliminary investigations on the levitation and thermal treatment of germania silicate gel-monoliths in the Pressure Facility Acoustic Levitator were done.

The digestion rate of dietary protein is a regulating factor for postprandial metabolism both in humans and animal models. However, few data exist about the habitual consumption of proteins with different digestion rates with regard to the development of body mass and diet-induced obesity. Here, we...... used a factorial ANOVA design to investigate the effects of protein form (intact vs. hydrolyzed casein) and protein level (16 vs. 32 energy percent protein) on body mass gain and adiposity in obesity-prone male C57BL/6J mice fed Western diets with 35 energy percent fat. Mice fed the hydrolyzed casein...... by hydrolyzed casein ingestion translated into decreased body and adipose tissue masses. We conclude that chronic consumption of extensively hydrolyzed casein reduces body mass gain and diet-induced obesity in male C57BL/6J mice....

The effects of heat treatment(heating temperature and pH) on the structures and emulsifying properties of caseins were systematically studied by spectroscopy.Heat treatment from 60 to 100 ℃ resulted in an increase in their fluorescence intensity,hydrodynamic diameter,turbidity and emulsifying activity index,but decreased the size polydispersity of caseins.In the pH range of 5.5 to 7.0,the fluorescence intensity,hydrodynamic diameter,turbidity and emulsifying properties decreased with increased heating pH,but the size polydispersity of caseins increased with increased pH.The relationship between the surface fluorescence intensity and emulsifying activity was also investigated,revealing a correlation coefficient of 0.90.These results suggested that heat treatment could be used to modify the structures and emulsifying properties of caseins by appropriately selecting heating conditions.

A reaction to casein, along with β-lactoglobulin, is a main cause of milk allergies, and also is a useful indicator of protein in allergic analyses. In the present study, a simple casein sensor was developed based on the interaction between a phosphate group of casein and electroactive [Ru(NH3)6](3+). We evaluated the voltammetric behavior of a casein-[Ru(NH3)6](3+) complex using a glassy carbon electrode. When the ruthenium(III) complex was combined with the phosphate groups of casein, the structure of the casein was changed. Since the hydrophobicity of casein was increased due to the binding, the casein was adsorbed onto the electrode. Furthermore, we modified an electrode with a ruthenium(III) ions/collagen film. When the sensor was applied to the detection of the casein contained in milk, the values coincided with those indicated by the manufacturer. Accordingly, this electrode could be a powerful sensor for the determination of casein in several foods.

of the Immune Response to Gluten and Casein in Autism Spectrum Disorders PRINCIPAL INVESTIGATOR: Armin Alaedini, PhD CONTRACTING... Gluten and Casein in Autism Spectrum Disorders 5a. CONTRACT NUMBER 5b. GRANT NUMBER W81XWH 10-1-0887 5c. PROGRAM ELEMENT NUMBER 6. AUTHOR(S...heterogeneity within this spectrum and the lack of biomarkers to characterize disease phenotypes and to understand treatment outcome. Dietary gluten

Full Text Available Have been determined the technical characteristics and elemental composition of milk casein. Pyrolysis experiments of casein carried out at different heating temperatures and determined the yields of obtained solid (biochar, liquid (tar and pyrolysis water and gas products. A temperature around 550ºC determined as an optimal heating temperature of pyrolysis and approximately 28.33% biochar, 37.38% tar, 13.23% pyrolysis water and 20.84% gas obtained after pyrolysis. First time a biochar with higher content of nitrogen was obtained by pyrolysis of casein and determined it is elemental composition and technical specifications. The porous structure of casein biochar was characterized by mercury porosimeter and SEM analysis confirmed that casein biochar has mostly meso and macro pores. The casein tar had the elemental composition: C-66.7%, H-8.3%, N-12.1%, O-12.9% and was completely soluble in 1-methyl-2-pyrroldinone. The tar consisted mostly of moderate molecular mass components with SEC elution times between 18-26 min and an estimated mass range up to 3000-5000 mass units as well as some larger size components, possibly 3-dimentional. The property and determined chemical composition of casein tar by GC/MS analysis were an evidence for using it as a curing agent for crosslinking reactions of epoxy resins. The necessary amount of tar for curing reaction of epoxy resin was determined experimentally as a 15-20% for the stoichiometric amount of reactive epoxy groups (15-20% epoxy group content in epoxy resin and obtained cured epoxy resin with 95% degree of crosslinking reaction. Have been suggested several curing reaction schemes of epoxy resin with amines, nitriles and phenols of the casein tar

Aim: Sheep milk-born bioactive peptides have been found to exhibit various physiological activities. The present work was designed with the aim to evaluate the effect of boiling on antidiabetic property of peptides derived from sheep milk caseinate on hydrolysis with three different proteases. Materials and Methods: In this investigation, casein prepared from raw and boiled sheep milk was hydrolyzed by three commercially available proteases (trypsin, pepsin, and chymotrypsin). These hydrol...

Pulse Field Gel Electrophoresis (PFGE) is a powerful genotyping technique used for the separation of large DNA molecules (entire genomic DNA) after digesting it with unique restriction enzymes and applying to a gel matrix under the electric field that periodically changes direction. PFGE is a variation of agarose gel electrophoresis that permits analysis of bacterial DNA fragments over an order of magnitude larger than that with conventional restriction enzyme analysis. It provides a good representation of the entire bacterial chromosome in a single gel with a highly reproducible restriction profile, providing clearly distinct and well-resolved DNA fragments.

This report describes work performed during the second year of the project, ''Conformance Improvement Using Gels.'' The project has two objectives. The first objective is to identify gel compositions and conditions that substantially reduce flow through fractures that allow direct channeling between wells, while leaving secondary fractures open so that high fluid injection and production rates can be maintained. The second objective is to optimize treatments in fractured production wells, where the gel must reduce permeability to water much more than that to oil. Pore-level images from X-ray computed microtomography were re-examined for Berea sandstone and porous polyethylene. This analysis suggests that oil penetration through gel-filled pores occurs by a gel-dehydration mechanism, rather than a gel-ripping mechanism. This finding helps to explain why aqueous gels can reduce permeability to water more than to oil. We analyzed a Cr(III)-acetate-HPAM gel treatment in a production well in the Arbuckle formation. The availability of accurate pressure data before, during, and after the treatment was critical for the analysis. After the gel treatment, water productivity was fairly constant at about 20% of the pre-treatment value. However, oil productivity was stimulated by a factor of 18 immediately after the treatment. During the six months after the treatment, oil productivity gradually decreased to approach the pre-treatment value. To explain this behavior, we proposed that the fracture area open to oil flow was increased substantially by the gel treatment, followed by a gradual closing of the fractures during subsequent production. For a conventional Cr(III)-acetate-HPAM gel, the delay between gelant preparation and injection into a fracture impacts the placement, leakoff, and permeability reduction behavior. Formulations placed as partially formed gels showed relatively low pressure gradients during placement, and yet substantially reduced the

Bovine beta-casein (beta-CN) is a highly amphiphilic micellising phospho-protein showing chaperone-like activity in vitro. Recently, existence of multiple sequential epitopes on beta-CN polypeptide chain in both hydrophilic-polar (psi) and hydrophobic-apolar domains (phi) has been evidenced. In order to clarify specific contribution of polar and apolar domains in micellisation process and in shaping immunoreactivity of beta-CN, its dimeric/bi-amphiphilic "quasi palindromic" forms covalently connected by a disulfide bond linking either N-terminal (C4 beta-CND) or C-terminal domain (C208 beta-CND) were produced and studied. Depending on the C- or N-terminal position of inserted cysteine, each dimeric beta-CN contains one polar/apolar region at the centre and two external hydrophobic/hydrophilic ends. Consequently, such casein dimers have radically different polarities/hydrophobicities on their outside surfaces. Dynamic light scattering (DLS) measurements indicate that these dimeric casein molecules form micelles of different sizes depending on arrangement of polar fragments of the beta-CN mutants in their constrained dimers. Non-aggregated dimers have different hydrodynamic diameters that could be explained by their different geometries. Measurements of fluorescence showed more hydrophobic environment of Trp residues of C208 beta-CND, while in similar experimental conditions Trp residues of C4 beta-CND and native beta-CN were more exposed to the polar medium. Both fluorescence and DLS studies showed greater propensity for micellisation of the dimeric beta-CNs, suggesting that the factors inducing the formation of micelles are stronger in the bi-amphiphilic dimers. 1-anilino-naphthalene-8-sulfonate (ANS) binding studies showed different binding of ANS by these dimers as well as different exposition of ANS binding (hydrophobic) regions in the micellar states. The differences in fluorescence resonance energy transfer (FRET) profiles of C4 beta-CND and C208 beta-CND can

The adsorption behaviour of mixtures of the proteins β-casein and hydrophobin at the hydrophilic solid-liquid surface have been studied by neutron reflectivity. The results of measurements from sequential adsorption and co-adsorption from solution are contrasted. The adsorption properties of protein mixtures are important for a wide range of applications. Because of competing factors the adsorption behaviour of protein mixtures at interfaces is often difficult to predict. This is particularly true for mixtures containing hydrophobin as hydrophobin possesses some unusual surface properties. At β-casein concentrations ⩾0.1wt% β-casein largely displaces a pre-adsorbed layer of hydrophobin at the interface, similar to that observed in hydrophobin-surfactant mixtures. In the composition and concentration range studied here for the co-adsorption of β-casein-hydrophobin mixtures the adsorption is dominated by the β-casein adsorption. The results provide an important insight into how the competitive adsorption in protein mixtures of hydrophobin and β-casein can impact upon the modification of solid surface properties and the potential for a wide range of colloid stabilisation applications.

We present GelTouch, a gel-based layer that can selectively transition between soft and stiff to provide tactile multi-touch feedback. It is flexible, transparent when not activated, and contains no mechanical, electromagnetic, or hydraulic components, resulting in a compact form factor (a 2mm thin...... touchscreen layer for our prototype). The activated areas can be morphed freely and continuously, without being limited to fixed, predefined shapes. GelTouch consists of a poly(N-isopropylacrylamide) gel layer which alters its viscoelasticity when activated by applying heat (>32 C). We present three different...... a tablet with 6x4 tactile areas, enabling a tactile numpad, slider, and thumbstick. We show that the gel is up to 25 times stiffer when activated and that users detect tactile features reliably (94.8%)....

Pancreatic cancer is one of the most common causes of cancer death in western civilization. The five-year survival rate is below 1% and of the 10% of patients with resectable disease only around one-fifth survives 5 years. Survival rates have not changed much during the last 20 years, demonstrating the inefficacy of current available therapies. To improve the prognosis of pancreatic cancer, there is the need to develop effective non-surgical treatment for this disease. The protein kinase casein kinase II (CK2) is a ubiquitously expressed serine-threonine kinase and its activity is enhanced in all human tumors examined so far. The contribution of CK2 to the tumor maintenance of pancreatic cancer has not been investigated. To investigate the function of CK2 in pancreatic cancer cells we used the CK2 specific inhibitors 5,6-Dichloro-1-beta-D-ribofuranosylbenzimidazole and Apigenin. Furthermore, we interfered with CK2 expression using CK2 specific siRNAs. Interfering with CK2 function led to a reduction of pancreatic cancer cell viability, which was due to caspase-dependent apoptosis. The induction of apoptosis correlated with a reduced NF-kappaB-dependent transcriptional activity. This study validates CK2 as a molecular drug target in a preclinical in vitro model of pancreatic cancer.

Although the immigration of water molecule, and diffusion and traversing of oxygen can be prevented by the edible film prepared through sodium caseinate, which plays a good protection role for the food, the strong hydrophilicity makes its watertightness and mechanical properties become inferior. Because the toughness and water resistance of SC films can be enhanced by glycerol (G) as an additive, it is necessary to elucidate the interaction between G and SC through the spectral characteristics such as fluorescence spectra, infrared spectra and UV spectra. The results show that the fluorescence intensity of SC decreases due to the addition of G. The binding constant obtained by the double logarithmic regression curve analysis is 1. 127 x 10(3) L . mol-1 and the number of binding sites reaches 1. 161. It indicates that the weak chemical bond is primary between G and SC molecules; From IR the absorption peaks of SC are almost the same before and after adding G. However, there is a certain difference among their absorption intensities. It reveals that the secondary structure of SC is affected, β folding length decreases, α helix, random coil structure, β angle structure increases, and the intermolecular hydrogen bond is strengthened; From UV the peptide bond structure of SC is not changed after the addition of G, but the polymer with larger molecular weight, which is formed by non-covalent bond, makes the peak intensity decrease. The research gives the mode of G and SC from the molecular level.

Replication of Mayaro virus in Vero cells induces dramatic cytopathic effects and cell death. In this study, we have evaluated the role of casein kinase 2 (CK2) during Mayaro virus infection cycle. We found that CK2 was activated during the initial stages of infection ( approximately 36% after 4h). This activation was further confirmed when the enzyme was partially purified from the cellular lysate either by Mono Q 5/5Hr column or heparin-agarose column. Using this later column, we found that the elution profile of CK2 activity from infected cells was different from that obtained for control cell enzyme, suggesting a structural modification of CK2 after infection. Treatment of infected cells with a cell-permeable inhibitor of CK2, dichloro-1-(beta-D-ribofuranosyl)benzimidazole (DRB), abolished the cytopathic effect in a dose-dependent manner. Together this set of data demonstrates for the first time that CK2 activity in host cells is required in Mayaro virus infection cycle.

Many nanoemulsions are currently formulated with synthetic surfactants. The objective of the present work was to study the possibility of blending sodium caseinate (NaCas) and lecithin to prepare transparent thyme oil nanoemulsions. Thyme oil was emulsified using NaCas and soy lecithin individually or in combination at neutral pH by shear homogenization. The surfactant combination improved the oil content in transparent/translucent nanoemulsions, from 1.0% to 2.5% w/v for 5% NaCas with and without 1% lecithin, respectively. Nanoemulsions prepared with the NaCas-lecithin blend had hydrodynamic diameters smaller than 100 nm and had significantly smaller and more narrowly distributed droplets than those prepared with NaCas or lecithin alone. Particle dimension and protein surface load data suggested the coadsorption of both surfactants on oil droplets. These characteristics of nanoemulsions minimized destabilization mechanisms of creaming, coalescence, and Ostwald ripening, as evidenced by no significant changes in appearance and particle dimension after 120-day storage at 21 °C.

Delivery systems prepared with natural biopolymers are of particular interests for applications in food, pharmaceutics and biomedicine. In this study, nanocomplex particles of sodium caseinate (NaCas) and pectin were fabricated and investigated as potential oral delivery vehicles. Nanocomplexes were prepared with three mass ratios of NaCas/pectin by acidification using glucono-δ-lactone and thermal treatment. NaCas/pectin at 1:1 mass ratio resulted in dispersions with the lowest turbidity and the smallest and most uniform nanocomplexes. Thermal treatment at 85 °C for 30 min facilitated the formation of stable, compact, and spherical nanocomplexes. Heating not only greatly increased the yield of nanocomplexes but also significantly improved the encapsulation capability of rutin studied as a model compound. Pectin in nanocomplexes delayed the hydrolysis of NaCas by pepsin at gastric conditions and enabled the controlled release of most rutin in simulated intestinal conditions. The nanocomplexes based on food-sourced biopolymers have promising features for oral delivery of nutrients and medicines.

目的 检测良性前列腺增生(BPH)患者组织中蛋白表达的特点,了解有无与BPH相关的特异性蛋白表达.方法 通过双向电泳方法 所发现的检测BPH组织特异性蛋白表达,αs1-Casein进行定量免疫组化研究来验证该蛋白表达的特异性.结果 通过对正常前列腺组织,BPH及前列腺癌组织进行免疫组织化学染色,发现αs1-Casein在90%(20/22)的BPH组织中呈弱到强表达,而在正常前列腺组织中未见表达(0/10),在前列腺癌组织中的表达也不足10%(3/30).结论 αs1-Casein可能是一种BPH诊断的生物标记物.%Objective To analyze the characteristic of protein profile of benign prostatic hyperplasia (BPH) tissue and to further find the special proteins associated with BPH. Methods Protein profiles of human expressed prostatic secretion (EPS) samples obtained from BPH patients and normal males were comparatively analyzed by two-dimensional gel electrophoresis. In the resulting electrophoretograms, several differentially expressed proteins were identified. In this study, a distinct expressed protein αs1-casein was validated in BPH. Results Immunohistochemistry analysis showed that αs1-casein was mainly distributed in the epithelium of hyperplastic prostate. The level of the exression of αs1-casein in the tissue of BPH was found to be higher than that in prostatic carcinoma; whereas no asl-casein was detected in normal tissue.Western blotting analysis was also revealed same expression pattern of asl-casein was detected in BPH-EPS and BPH-free EPS. Conclusion asl-casein might be associated with BPH development and be used as a potential biological marker for the diagnosis of BPH.

The aim of this study was to evaluate the efficacy of tooth mousse containing 10% casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) in reducing tooth sensitivity associated with in-office vital tooth whitening. In-office tooth whitening was performed for 51 participants using 35% hydrogen peroxide gel in a single visit. After the procedure, each participant was randomly assigned to one of three groups: gel without desensitizing agent (n=17), gel with 2% sodium fluoride (n=17), gel with 10% CPP-ACP (n=17). A small amount of the desensitizing gel assigned for each participant was applied directly on the labial surfaces of teeth and left undisturbed for three minutes. The participants were asked to apply the gel assigned to them for three minutes twice daily after brushing their teeth, and to continue this for 14 days. The participants were asked to return for follow-up visits after 24 hours and on days 3, 7, and 14, at which time teeth shade changes were assessed by one evaluator using a value-oriented Vita classic shade guide. The incidence, duration, and intensity of tooth sensitivity experienced was self-assessed on a daily basis for the 14-day study period using a visual analog scale (VAS). The effect of the three gels on tooth sensitivity was assessed using one-way analysis of variance and a χ (2) test (α=0.05). The general linear model was used to compare intensity-level differences in the three studied groups and for shade stability over the follow-up period. The results of this study showed that all three gels decreased the intensity of sensitivity associated with tooth whitening. The intensity of sensitivity was lower in the fluoride group than in the other two groups; however, it was not statistically significant (p=0.112 and p=0.532 on day 1 and day 2, respectively). The average shade change was 6.8. None of the tested materials affected the efficacy of tooth whitening, but the shade change among the fluoride group showed more color stability

The analysis of casein polymorphisms was carried out in West Africa goat populations: Red Sokoto (n = 57), West African Dwarf Nigeria (n = 27), West African Dwarf Cameroon (n = 39), and Borno (n = 37). The 4 casein genes alpha(s1) (CSN1S1), beta (CSN2), alpha(s2) (CSN1S2), and kappa (CSN3) were typed at the DNA level. No null alleles were found in any of the genes analyzed. A PCR single-strand conformation polymorphism method was implemented for the identification of CSN1S1*F allele simultaneously with A/0(1), B/E, N and the new allele. The allele differed from CSN1S1*B by a synonymous transversion TCG-->TCT in the codon corresponding to Ser(66) of the mature protein. The new allele, named CSN1S1*B', occurred at a high frequency in all the populations, ranging from 0.295 (West African Dwarf Cameroon) to 0.405 (Borno). A greater frequency was found for alleles associated with high alpha(s1)-casein quantity, as has already been observed in the goat populations from the Mediterranean area. The intermediate E allele occurred only in the Red Sokoto and at a low frequency. The faint F allele occurred in 3 populations at frequencies lower than 0.03. Linkage disequilibrium occurred in all the populations, with highly significant differences in Borno, Red Sokoto, and West Africa Dwarf Nigeria, and significant differences in West Africa Dwarf Cameroon. Only 10 haplotypes showed frequencies > or =0.05 in at least 1 of the 4 populations considered, and the overall frequency was >0.1 only for 4 haplotypes: BAAB, B'ACA, ACAB, and BACA (in the order CSN1S1-CSN2-CSN1S2-CSN3). Haplotype BAAB, postulated as an ancestral haplotype in previous studies, was the most common haplotype in all breeds except Borno, where B'ACA was predominant. The results obtained are of considerable significance given that very little information exists on the subject for African goats. The high frequency of strong alleles in the calcium-sensitive caseins as well as the high linkage disequilibrium found

Polymorphism of goat casein gene closely linked to the quality of milk protein. κ-casein is one of the casein fractions that influence the shape and stability of grain milk. This study is aimed to identify the variation of genotype κ-casein and related with milk quality from Etawah grade, Saanen and their crossbreed (PE-SA). The number of dairy goats used in this study was 150 animals consisted of Etawah grade (48 animals), Saanen (51 animals) and PE-SA (51 animals). Steps of experiment were:...

Low-molecular-weight (LMW) emulsifiers are used to promote controlled destabilization in many dairy-type emulsions in order to obtain stable foams in whippable products. The relation between fat globule aggregation induced by three LMW emulsifiers, lactic acid ester of monoglyceride (LACTEM), saturated monoglyceride (GMS), and unsaturated monoglyceride (GMU) and their effect on interfacial protein displacement was investigated. It was found that protein displacement by LMW emulsifiers was not necessary for fat globule aggregation in emulsions, and conversely fat globule aggregation was not necessarily accompanied by protein displacement. The three LMW emulsifiers had very different effects on emulsions. LACTEM induced shear instability of emulsions, which was accompanied by protein displacement. High stability was characteristic for emulsions with GMS where protein was displaced from the interface. Emulsions containing GMU were semisolid, but only low concentrations of protein were detected in the separated serum phase. The effects of LACTEM, GMS, and GMU may be explained by three different mechanisms involving formation of interfacial α-gel, pickering stabilization and increased exposure of bound casein to the water phase. The latter may facilitate partial coalescence. Stabilizing hydrocolloids did not have any effect on the LMW emulsifiers' ability to induce protein displacement.

Although in-office bleaching has been proven successful for bleaching teeth, controversy exists from morphological alterations in enamel morphology due to mineral loss and tooth sensitivity. This preliminary study aimed to evaluate the efficacy of a novel in-office tooth bleaching technique modified with a casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) paste (MI paste-MI) and its effect on the enamel morphology and tooth sensitivity. Three patients received a 35% hydrogen peroxide (Whiteness HP-HP) dental bleaching system. HP was prepared and applied on the teeth on one of the hemiarches, whilst teeth on the other hemiarch were bleached with a mixture of HP and MI. Tooth color, epoxy resin replicas, and sensitivity levels were evaluated in the upper incisors. The results were analyzed descriptively. Right and left incisors showed similar color change after bleaching. Incisors bleached with the mixture of HP and MI presented unaltered enamel surfaces and lower sensitivity levels. The currently tested tooth bleaching technique did not reduce the gel effectiveness while decreasing hypersensitivity levels and protecting the enamel against surface alterations caused by the high-concentration bleaching peroxide tested. The concomitant use of MI Paste and high-concentration hydrogen peroxide might be a successful method for decreasing tooth sensitivity and limiting changes in the enamel morphology during in-office bleaching.

In adults, dietary protein seems to induce weight loss and dairy proteins may be insulinotropic. However, the effect of milk proteins in adolescents is unclear. The objective was to test whether milk and milk proteins reduce body weight, waist circumference, homeostatic model assessment, plasma insulin, and insulin secretion estimated as the plasma C-peptide concentration in overweight adolescents. Overweight adolescents (n = 203) aged 12-15 y with a BMI of 25.4 ± 2.3 kg/m(2) (mean ± SD) were randomized to 1 L/d of skim milk, whey, casein, or water for 12 wk. All milk drinks contained 35 g protein/L. Before randomization, a subgroup of adolescents (n = 32) was studied for 12 wk before the intervention began as a pretest control group. The effects of the milk-based test drinks were compared with baseline (wk 0), the water group, and the pretest control group. Diet and physical activity were registered. Outcomes were BMI-for-age Z-scores (BAZs), waist circumference, plasma insulin, homeostatic model assessment, and plasma C-peptide. We found no change in BAZ in the pretest control and water groups, whereas it was greater at 12 wk in the skim milk, whey, and casein groups compared with baseline and with the water and pretest control groups. The plasma C-peptide concentration increased from baseline to wk 12 in the whey and casein groups and increments were greater than in the pretest control (P milk or water group. These data suggest that high intakes of skim milk, whey, and casein increase BAZs in overweight adolescents and that whey and casein increase insulin secretion. Whether the effect on body weight is primary or secondary to the increased insulin secretion remains to be elucidated.

Wnt signaling is critical to many aspects of development, and aberrant activation of the Wnt signaling pathway can cause cancer. Dishevelled (Dvl) protein plays a central role in this pathway by transducing the signal from the Wnt receptor complex to the beta-catenin destruction complex. Dvl also plays a pivotal role in the planar cell polarity pathway that involves the c-Jun N-terminal kinase (JNK). How functions of Dvl are regulated in these two distinct pathways is not clear. We show that deleting the C-terminal two-thirds of Dvl, which includes the PDZ and DEP domains and is essential for Dvl-induced JNK activation, rendered the molecule a much more potent activator of the beta-catenin pathway. We also found that casein kinase Iepsilon (CKIepsilon), a previously identified positive regulator of Wnt signaling, stimulated Dvl activity in the Wnt pathway, but dramatically inhibited Dvl activity in the JNK pathway. Consistent with this, overexpression of CKIepsilon in Drosophila melanogaster stimulated Wnt signaling and disrupted planar cell polarity. We also observed a correlation between the localization and the signaling activity of Dvl in the beta-catenin pathway and the JNK pathway. Furthermore, by using RNA interference, we demonstrate that the Drosophila CKIepsilon homologue Double time positively regulates the beta-catenin pathway through Dvl and negatively regulates the Dvl-induced JNK pathway. We suggest that CKIepsilon functions as a molecular switch to direct Dvl from the JNK pathway to the beta-catenin pathway, possibly by altering the conformation of the C terminus of Dvl.

Consumption of cow's milk is recommended in many countries. Observational and intervention studies show that cow's milk most likely has a positive influence on growth in children. The strongest evidence comes from observational studies and intervention studies in low-income countries, but there are also observational studies from high-income countries showing positive associations between milk intake and growth. Milk seems thus to have a specific stimulating effect on linear growth, not only in developing countries with high rates of malnutrition, but also in industrialized countries. However, it is not known which components in milk stimulate growth. Possible components are proteins, minerals, vitamins or combinations of these. Cow's milk proteins have a high protein quality, and whey has a slightly higher quality than casein, according to some indices based on amino acid composition. Studies, mainly from sport medicine, have suggested that whey protein also has the potential to increase muscle mass. Whether whey improves body composition to a larger extent than other milk proteins is not clear. The mechanism behind a possible growth-stimulating effect of milk and milk components is likely to be through a stimulation of insulin-like growth factor-I synthesis and maybe insulin secretion. In conclusion, there is strong evidence that milk stimulates linear growth. The mechanism is not yet clear, and more intervention studies are needed to understand which components in milk are responsible for the growth stimulation. The effects of milk on linear growth and adult height may have both positive and negative long-term implications.

Methods have been devised for detecting polymorphisms in the bovine beta- and kappa-casein genes using the polymerase chain reaction (PCR) followed either by restriction enzyme digestion (to reveal a restriction fragment length polymorphism (RFLP] or by hybridization of an allele-specific oligonucleotide. These methods, as well as being faster and more sensitive than traditional RFLP methods, are of more general applicability since they can detect any change in DNA sequence. They require only a small sample of blood or semen and are applicable to animals of any age or sex. These methods make possible large-scale screening and thus selection for alleles at these loci. Typing of blood DNA can give erroneous results when the animal concerned is a twin; however, this can be overcome by retesting using milk or semen. Analysis of the kappa-casein genotype of Holstein-Friesian bulls gives frequencies for the A and B alleles of 0.80 and 0.20 respectively. Selection in favour of the B allele, which is superior for cheese production, could thus have a large effect. The A3 and B alleles at the beta-casein locus have been shown to be rare in the Holstein-Friesian population. Linkage disequilibrium exists between beta-casein B and kappa-casein B.

The effects of lactose on the changes in the composition and size of casein micelles induced by high-pressure treatment and the related mechanism of action were investigated. Dispersions of ultracentrifuged casein micelle pellets with 0-10% (w/v) lactose were subjected to high pressure (400 MPa) at 20 °C for 40 min. The results indicated that the level of non-sedimentable caseins was positively related to the amount of lactose added prior to pressure treatment, and negatively correlated to the size. A mechanism for the pressure-induced, lactose-dependent changes in the casein micelles is proposed. Lactose inhibits the hydrophobic interactions between the micellar fragments during or after pressure release, through the hydrophilic layer formed by their hydrogen bonds around the micellar fragments. In addition, lactose does not favour the association between calcium and the casein aggregates after pressure release. Due to these two functions, lactose inhibited the formation of larger micelles after pressure treatment.

Although lignocellulosic materials have a good potential to substitute current feedstocks used for ethanol production, conversion of these materials to fermentable sugars is still not economical through enzymatic hydrolysis. High cost of cellulase has prompted research to explore techniques that can prevent from enzyme deactivation. Colloidal proteins of casein can form monolayers on hydrophobic surfaces that alleviate the de-activation of protein of interest. Scanning electron microscope (SEM), fourier transform infrared spectroscopy (FT-IR), capillary electrophoresis (CE), and Kjeldahl and BSA protein assays were used to investigate the unknown mechanism of action of induced cellulase activity during hydrolysis of casein-treated biomass. Adsorption of casein to biomass was observed with all of the analytical techniques used and varied depending on the pretreatment techniques of biomass. FT-IR analysis of amides I and II suggested that the substructure of protein from casein or skim milk were deformed at the time of contact with biomass. With no additive, the majority of one of the cellulase mono-component, 97.1 ± 1.1, was adsorbed to CS within 24 h, this adsorption was irreversible and increased by 2% after 72 h. However, biomass treatment with skim-milk and casein reduced the adsorption to 32.9% ± 6.0 and 82.8% ± 6.0, respectively. PMID:23118515

Full Text Available Although lignocellulosic materials have a good potential to substitute current feedstocks used for ethanol production, conversion of these materials to fermentable sugars is still not economical through enzymatic hydrolysis. High cost of cellulase has prompted research to explore techniques that can prevent from enzyme deactivation. Colloidal proteins of casein can form monolayers on hydrophobic surfaces that alleviate the de-activation of protein of interest. Scanning electron microscope (SEM, fourier transform infrared spectroscopy (FT-IR, capillary electrophoresis (CE, and Kjeldahl and BSA protein assays were used to investigate the unknown mechanism of action of induced cellulase activity during hydrolysis of casein-treated biomass. Adsorption of casein to biomass was observed with all of the analytical techniques used and varied depending on the pretreatment techniques of biomass. FT-IR analysis of amides I and II suggested that the substructure of protein from casein or skim milk were deformed at the time of contact with biomass. With no additive, the majority of one of the cellulase mono-component, 97.1 ± 1.1, was adsorbed to CS within 24 h, this adsorption was irreversible and increased by 2% after 72 h. However, biomass treatment with skim-milk and casein reduced the adsorption to 32.9% ± 6.0 and 82.8% ± 6.0, respectively.

By combining the osmotic stress technique with small-angle x-ray scattering measurements, we followed the structural response of the casein micelle to an overall increase in concentration. When the aqueous phase that separates the micelles is extracted, they behave as polydisperse repelling spheres and their internal structure is not affected. When they are compressed, the micelles lose water and shrink to a smaller volume. Our results indicate that this compression is nonaffine, i.e., some parts of the micelle collapse, whereas other parts resist deformation. We suggest that this behavior is consistent with a spongelike casein micelle having a triple hierarchical structure. The lowest level of the structure consists of the CaP nanoclusters that serve as anchors for the casein molecules. The intermediate level consists of 10- to 40-nm hard regions that resist compression and contain the nanoclusters. Those regions are connected and/or partially merged with each other, thus forming a continuous and porous material. The third level of structure is the casein micelle itself, with an average size of 100 nm. In our view, such a structure is consistent with the observation of 10- to 20-nm casein particles in the Golgi vesicles of lactating cells: upon aggregation, those particles would rearrange, fuse, and/or swell to form the spongelike micelle.

The aim of this study was to assess biologically safer components as alternatives to egg yolk for the frozen storage of ram semen using casein, coconut or palm oil in either Salamon's diluent (S) or a swim-up medium (SU). Ejaculates were frozen as pellets and sperm motility (subjectively) and acrosome integrity (FITC-PNA/PI) by flow cytometry were assessed at 0, 3 and 6h after thawing and incubation at 37°C. Three experiments were done: different concentrations of palm oil (5%, 10% and 20%); casein added as emulsifier and protective agent; and differences between egg yolk, coconut and palm oil in S and SU. 20% of oil added to SU accounted for a lesser percentage (Poil levels on viable cells. When casein was added to diluents containing 5% of palm oil, no differences were found between palm or casein (P>0.05). No differences were found when S and SU were compared neither as groups nor between S alone and containing coconut or palm oil; however, SU alone yielded less motility than SU 5% coconut. However, in both groups, S and SU, egg yolk accounted for the greatest values in both bases. These results indicate that none of biologically safer media components (casein, palm or coconut oil) used in this study maintained the function of ram spermatozoa after freeze-thawing better than S-containing egg yolk. The application of vegetable oils as substitutes for egg yolk in diluents for the cryopreservation of ram spermatozoa requires further research.

Mechanical triggering of gelation of an organic solution by a carbazole-based bisurea organogelator is described. Both the duration of the mechanical stimulation and the gelator concentration control the gelation process and the characteristics of the gel obtained.

Full Text Available Caseins, the main milk proteins, interact with colloidal calcium phosphate to form the casein micelle. The mesostructure of this supramolecular assembly markedly influences its nutritional and technological functionalities. However, its detailed molecular organization and the cellular mechanisms involved in its biogenesis have been only partially established. There is a growing body of evidence to support the concept that α(s1-casein takes center stage in casein micelle building and transport in the secretory pathway of mammary epithelial cells. Here we have investigated the membrane-associated form of α(s1-casein in rat mammary epithelial cells. Using metabolic labelling we show that α(s1-casein becomes associated with membranes at the level of the endoplasmic reticulum, with no subsequent increase at the level of the Golgi apparatus. From morphological and biochemical data, it appears that caseins are in a tight relationship with membranes throughout the secretory pathway. On the other hand, we have observed that the membrane-associated form of α(s1-casein co-purified with detergent-resistant membranes. It was poorly solubilised by Tween 20, partially insoluble in Lubrol WX, and substantially insoluble in Triton X-100. Finally, we found that cholesterol depletion results in the release of the membrane-associated form of α(s1-casein. These experiments reveal that the insolubility of α(s1-casein reflects its partial association with a cholesterol-rich detergent-resistant microdomain. We propose that the membrane-associated form of α(s1-casein interacts with the lipid microdomain, or lipid raft, that forms within the membranes of the endoplasmic reticulum, for efficient forward transport and sorting in the secretory pathway of mammary epithelial cells.

This research project had two objectives. The first objective was to identify gel compositions and conditions that substantially reduce flow through fractures that allow direct channeling between wells, while leaving secondary fractures open so that high fluid injection and production rates can be maintained. The second objective was to optimize treatments in fractured production wells, where the gel must reduce permeability to water much more than that to oil.

Among the various crystallization techniques, crystallization in gels has found wide applications in the fields of biomineralization and macromolecular crystallization in addition to crystallizing materials having nonlinear optical, ferroelectric, ferromagnetic, and other properties. Furthermore, by using this method it is possible to grow single crystals with very high perfection that are difficult to grow by other techniques. The gel method of crystallization provides an ideal technique to study crystal deposition diseases, which could lead to better understanding of their etiology. This chapter focuses on crystallization in gels of compounds that are responsible for crystal deposition diseases. The introduction is followed by a description of the various gels used, the mechanism of gelling, and the fascinating phenomenon of Liesegang ring formation, along with various gel growth techniques. The importance and scope of study on crystal deposition diseases and the need for crystal growth experiments using gel media are stressed. The various crystal deposition diseases, viz. (1) urolithiasis, (2) gout or arthritis, (3) cholelithiasis and atherosclerosis, and (4) pancreatitis and details regarding the constituents of the crystal deposits responsible for the pathological mineralization are discussed. Brief accounts of the theories of the formation of urinary stones and gallstones and the role of trace elements in urinary stone formation are also given. The crystallization in gels of (1) the urinary stone constituents, viz. calcium oxalate, calcium phosphates, uric acid, cystine, etc., (2) the constituents of the gallstones, viz. cholesterol, calcium carbonate, etc., (3) the major constituent of the pancreatic calculi, viz., calcium carbonate, and (4) cholic acid, a steroidal hormone are presented. The effect of various organic and inorganic ions, trace elements, and extracts from cereals, herbs, and fruits on the crystallization of major urinary stone and gallstone

Full Text Available Aerogel conditioning of the chitosan makes it possible to prepare porous solids of significant specific surface. The increase in the chitosan concentration or the degree of acetylation decreases the specific surface of the synthesized chitosan gel. Whereas drying with supercritical CO2 more effectively makes it possible to preserve the volume of the spheres of gel and to have a more significant specific surface in comparison with evaporative drying.

This report describes work performed during the third and final year of the project, ''Conformance Improvement Using Gels.'' Corefloods revealed throughput dependencies of permeability reduction by polymers and gels that were much more prolonged during oil flow than water flow. This behavior was explained using simple mobility ratio arguments. A model was developed that quantitatively fits the results and predicts ''clean up'' times for oil productivity when production wells are returned to service after application of a polymer or gel treatment. X-ray computed microtomography studies of gels in strongly water-wet Berea sandstone and strongly oil-wet porous polyethylene suggested that oil penetration through gel-filled pores occurs by a gel-dehydration mechanism, rather than gel-ripping or gel-displacement mechanisms. In contrast, analysis of data from the University of Kansas suggests that the gel-ripping or displacement mechanisms are more important in more permeable, strongly water-wet sandpacks. These findings help to explain why aqueous gels can reduce permeability to water more than to oil under different conditions. Since cement is the most commonly used material for water shutoff, we considered when gels are preferred over cements. Our analysis and experimental results indicated that cement cannot be expected to completely fill (top to bottom) a vertical fracture of any width, except near the wellbore. For vertical fractures with apertures less than 4 mm, the cement slurry will simply not penetrate very far into the fracture. For vertical fractures with apertures greater than 4 mm, the slurry may penetrate a substantial distance into the bottom part of the fracture. However, except near the wellbore, the upper part of the fracture will remain open due to gravity segregation. We compared various approaches to plugging fractures using gels, including (1) varying polymer content, (2) varying placement (extrusion) rate

Edible films based on sodium caseinate and different saturated fatty acids, oleic acid, or beeswax were formulated. Film-forming emulsions were characterized in terms of particle size distribution, rheological behavior and surface tension. In order to evaluate the influence of lipids on sodium caseinate matrices, mechanical, optical, and water vapor barrier properties were studied, taking into account the effect of water content and film structure on such properties. Saturated fatty acids affected the film properties in a particular way due to the formation of bilayer structures which limited water vapor permeability, giving rise to nonflexible and more opaque films. Oleic acid and beeswax were less effective as water vapor barriers, although the former imparted more flexibility to the caseinate films and did not reduce the film transparency notably.

Agaricus bisporus and equine cytochrome c. Only for the first target β-casein was acting as a molecular chaperone i.e. its presence resulted in higher residual activity (higher degree of the function preservation). β-Casein did not have any influence on the residual activity of tyrosinase. Surprisingly...

Full Text Available Normal 0 21 false false false SK X-NONE X-NONE MicrosoftInternetExplorer4 The goal of work was identification of b - casein gene polymorphism in different breeds of cow. The beta - casein constitutes up to 45 % of the casein of bovine milk. The most common forms of beta-casein in dairy cattle breeds are A1 and A2, while B is less common. The b-casein A1 variant was associated with the incidence of diabetes mellitus 1st type, coronary heart disease and autism. The A2 variant reduces serum cholesterol. The material involved 287 cows (Simmental breed – 111 cows, Pinzgau breed – 89 cows, Holstein breed – 87 cows. Bovine genomic DNA was extracted from whole blood by using commercial kit and used in order to estimate b - casein genotypes by means of PCR-RFLP method. In the populations included in the study were detected all three genotypes – homozygote genotype A1A1, heterozygote genotype A1A2 and homozygote genotype A2A2 with frequencies 0.1261, 0.3333 and 0.5405 in Simmental breed; 0.1379, 0.4598 and 0.4023 in Holstein breed, 0.3034, 0.5168 and 0.1798 in Pinzgau breed. In population of Simmental breed and Holstein breed was higher frequency of allele A2 (0.7072 and 0.6322. In opposite, in population of Pinzgau breed was present higher frequency of the allele A1 (0.5618.

Two acidic proteins from 80-S ribosomes were isolated and purified to homogeneity. The purified acidic proteins could be phosphorylated by casein kinase using [gamma-32P]ATP and [gamma-32P]GTP as a phosphoryl donor. The proteins became phosphorylated in situ, too. Sodium dodecyl sulfate polyacryl......Two acidic proteins from 80-S ribosomes were isolated and purified to homogeneity. The purified acidic proteins could be phosphorylated by casein kinase using [gamma-32P]ATP and [gamma-32P]GTP as a phosphoryl donor. The proteins became phosphorylated in situ, too. Sodium dodecyl sulfate...

Serotonin (5-hydroxytriptamine, 5-HT) has an important role in milk volume homeostasis within the mammary gland during lactation. We have previously shown that the expression of β-casein, a differentiation marker in mammary epithelial cells, is suppressed via 5-HT-mediated inhibition of signal transduction and activator of transcription 5 (STAT5) phosphorylation in the human mammary epithelial MCF-12A cell line. In addition, the reduction of β-casein in turn was associated with 5-HT7 receptor expression in the cells. The objective of this study was to determine the mechanisms underlying the 5-HT-mediated suppression of β-casein and STAT5 phosphorylation. The β-casein level and phosphorylated STAT5 (pSTAT5)/STAT5 ratio in the cells co-treated with 5-HT and a protein kinase A (PKA) inhibitor (KT5720) were significantly higher than those of cells treated with 5-HT alone. Exposure to 100 μM db-cAMP for 6 h significantly decreased the protein levels of β-casein and pSTAT5 and the pSTAT5/STAT5 ratio, and significantly increased PTP1B protein levels. In the cells co-treated with 5-HT and an extracellular signal-regulated kinase1/2 (ERK) inhibitor (FR180294) or Akt inhibitor (124005), the β-casein level and pSTAT5/STAT5 ratio were equal to those of cells treated with 5-HT alone. Treatment with 5-HT significantly induced PTP1B protein levels, whereas its increase was inhibited by KT5720. In addition, the PTP1B inhibitor sc-222227 increased the expression levels of β-casein and the pSTAT5/STAT5 ratio. Our observations indicate that PTP1B directly regulates STAT5 phosphorylation and that its activation via the cAMP/PKA pathway downstream of the 5-HT7 receptor is involved in the suppression of β-casein expression in MCF-12A cells.

The digestion rate of dietary protein is a regulating factor for postprandial metabolism both in humans and animal models. However, few data exist about the habitual consumption of proteins with different digestion rates with regard to the development of body mass and diet-induced obesity. Here, ...... by hydrolyzed casein ingestion translated into decreased body and adipose tissue masses. We conclude that chronic consumption of extensively hydrolyzed casein reduces body mass gain and diet-induced obesity in male C57BL/6J mice....

Full Text Available In this paper, we introduce autonomous gel actuators driven by chemical energy. The polymer gels prepared here have cyclic chemical reaction networks. With a cyclic reaction, the polymer gels generate periodical motion. The periodic motion of the gel is produced by the chemical energy of the oscillatory Belouzov-Zhabotinsky (BZ reaction. We have succeeded in making synthetic polymer gel move autonomously like a living organism. This experimental fact represents the great possibility of the chemical robot.

-casein on the enzymatic activity of three targets was tested by monitoring enzyme activity after heat treatment and by measuring the intensity of scattered light during and after heat treatment. β-Casein was shown to interact at elevated temperatures with three selected targets:horseradish peroxidase, tyrosinase from...... Agaricus bisporus and equine cytochrome c. Only for the first target β-casein was acting as a molecular chaperone i.e. its presence resulted in higher residual activity (higher degree of the function preservation). β-Casein did not have any influence on the residual activity of tyrosinase. Surprisingly......, peroxidase activity of cytochrome c was increasing after heat treatment with β-casein (up to 518±9%). This indicates loss of the cytochrome c native conformation. Presence of bovine serum albumin (BSA) during heat treatment did not affect residual activity of tyrosinase and cytochrome c. Surprisingly...

followed by 30 h of enteral feeding with whey [protein fraction of milk formula based on whey (WHEY); n = 11] or casein and/or whey [protein fraction of milk formula based on a combination of casein and whey (CASEIN); n = 9]-based formulas. Sugar absorptive function was investigated at 6 and 30 h after......Preterm infants are susceptible to necrotizing enterocolitis (NEC). Using preterm pigs, we determined whether a whey–casein-based formula would be superior to a formula based on whey protein alone. Twenty cesarean-derived preterm pigs (92% gestation) were given total parenteral nutrition for 36 h...... studied in gut contents. Severity of NEC lesions was similar between diet groups but galactose absorption was markedly higher in CASEIN than in WHEY (P

In this work, a double-probe based immunoassay was developed for rapid and sensitive determination of β-casein in bovine milk samples. In the method, magnetic beads (MBs), employed as supports for the immobilization of anti-β-casein polyclonal antibody (PAb), were used as the capture probe. Colloidal gold nanoparticles (AuNPs), employed as a bridge for loading anti-β-casein monoclonal antibody (McAb) and horseradish peroxidase (HRP), were used as the amplification probe. The presence of β-casein causes the sandwich structures of MBs-PAb-β-casein-McAb-AuNPs through the interaction between β-casein and the anti-β-casein antibodies. The HRP, used as an enzymatic-amplified tracer, can catalytically oxidize the substrate 3,3',5,5'-tetramethylbenzidine (TMB), generating optical signals that are proportional to the quantity of β-casein. The linear range of the immunoassay was from 6.5 to 1520ngmL(-1). The limit of detection (LOD) was 4.8ngmL(-1) which was 700 times lower than that of MBs-antibody-HRP based immunoassay and 6-7 times lower than that from the microplate-antibody-HRP based assay. The recoveries of β-casein from bovine milk samples were from 95.0% to 104.3% that had a good correlation coefficient (R(2)=0.9956) with those obtained by an official standard Kjeldahl method. For higher sensitivity, simple sample pretreatment and shorter time requirement of the antigen-antibody reaction, the developed immunoassay demonstrated the viability for detection of β-casein in bovine milk samples.

Gel electrolytes, especially gel electrolytes for electrochemical cells, are generally described. In some embodiments, the gel electrolyte layers comprise components a) to c). Component a) may be at least one layer of at least one polymer comprising polymerized units of: a1) at least one monomer containing an ethylenically unsaturated unit and an amido group and a2) at least one crosslinker. Component b) may be at least one conducting salt and component c) may be at least one solvent. Electrodes may comprise the components a), d) and e), wherein component a) may be at least one layer of at least one polymer as described herein. Component d) may be at least one electroactive layer and component e) may be at least one ceramic layer. Furthermore, electrochemical cells comprising component a) which may be at least one layer of at least one polymer as described herein, are also provided.

Full Text Available Many kinds of stimuli-responsive polymer and gels have been developed and applied to biomimetic actuators or artificial muscles. Electroactive polymers that change shape when stimulated electrically seem to be particularly promising. In all cases, however, the mechanical motion is driven by external stimuli, for example, reversing the direction of electric field. On the other hand, many living organisms can generate an autonomous motion without external driving stimuli like self-beating of heart muscles. Here we show a novel biomimetic gel actuator that can walk spontaneously with a wormlike motion without switching of external stimuli. The self-oscillating motion is produced by dissipating chemical energy of oscillating reaction. Although the gel is completely composed of synthetic polymer, it shows autonomous motion as if it were alive.

Human casein kinase 2 alpha gene (CK-2-alpha) sequences have been localized within the human genome by in situ hybridization and somatic cell hybrid analysis using a CK-2 alpha cDNA as a probe. By in situ hybridization, the CK-2 alpha cDNA could be assigned to two different loci, one on 11p15.1-ter...

before exercise (CasPre), caseinate intake immediately after exercise (CasPost), whey intake immediately after exercise (Whey), or intake of a non-caloric control drink (Control). Muscle myofibrillar and collagen fractional synthesis rates (FSR) were measured by a primed continuous infusion of L-[1...... protein synthesis (MPS) to intakes of whey and caseinate after heavy resistance exercise in healthy elderly individuals, and, furthermore, to compare the timing effect of caseinate intake. Twenty-four elderly men and women (mean ± SEM; 68 ± 1 years) were randomized to one of four groups: caseinate intake......-(13) C]leucine using labeled proteins during a 6-h recovery period. No differences were observed in muscle myofibrillar and collagen FSR with Whey (0.09 ± 0.01%/h) compared with CasPost (0.09 ± 0.003%/h), and it did not differ between CasPre (0.10 ± 0.01%/h) and CasPost. MPS does not differ with whey...

Foam stability of casein micelle dispersions (CMDs) strongly depends on aggregate size. To elucidate the underlying mechanism, the role of interfacial and thin film properties was investigated. CMDs were prepared at 4 °C and 20 °C, designated as CMD4 °C and CMD20 °C. At equal protein concentrations,

We have investigated phosphorylation of human nucleoside diphosphate kinase (NDPK) and of homologous NDPK from different species by human casein kinase 2 (CK-2). The human NDPK isotypes A and B were phosphorylated by CK-2 in vitro both when the purified proteins and total lysate of HL-60 leukemia...

A model system of bovine colostrum and piscidin, a fish-derived antimicrobial peptide, was developed to study potential interactions of antimicrobial peptides in colostrum. We did not detect any antimicrobial activity of colostrum using the radial plate diffusion assay; in fact colostrum completely abrogated activity of added piscidin. This could not be explained by degradation of piscidin by colostrum, which was less than ten percent. We found that colostrum even protected piscidin against degradation by added proteases. We further observed that colostrum and milk rapidly quenched the fluorescence of fluorescein-piscidin but not that of fluorescein. This effect was not seen with BSA and the specific quenching of fluorescein-piscidin by colostrum was saturably inhibited with unlabeled piscidin. Size exclusion chromatography indicated that fluorescein-piscidin bound to casein micelles with no apparent binding to IgG or whey proteins. Further, addition of pure caseins was able to quench fluorescence of fluorescein-piscidin and to inhibit the antimicrobial activity of piscidin. The interaction between caseins and piscidin could be dissociated by guanidine hydrochloride and recovered piscidin had antimicrobial activity against bacteria. Based on our results we propose that caseins could be carriers for antimicrobial peptides in colostrum and milk.

Numerous peptides obtained by enzymatic digestion of food proteins have been reported to exhibit biological activities. In this study, the focus was placed on peptides of beta-casein from bovine milk after a gastro-analogous in vitro digestion with pepsin, a protease with broad specificity. In order to study the time course of the digestion, the process was stopped after specific times and the samples were subjected to HPLC separation followed by matrix-assisted laser desorption/ionization (MALDI) time-of-flight (TOF) and nanoelectrospray (nanoESI) quadrupole time-of-flight (qTOF) mass spectrometry. A combined sequencing approach using de novo interpretation and databases was employed. Overall, 100% of the beta-casein sequence was covered by identifying 125 peptides of 4-84 residues in length, including 3 phosphorylated species. The results show that the peptic hydrolysis starts at the C-terminus of the protein. The release of known bioactive peptides from beta-casein following the peptic digestion under simulated gastric conditions is unlikely with a few exceptions. Furthermore, an amino acid variation was found, providing evidence for the existence of an additional genetic variant of beta-casein.

Milk is an excellent source of bioactive peptides. However, the composition of the native milk peptidome has only been partially elucidated. The present study applied matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) directly or after prefractionation of the milk peptides by reverse-phase high-performance liquid chromatography (RP-HPLC) or OFFGEL fractionation for the comprehensive analysis of the peptide profile of raw milk. The peptide sequences were determined by MALDI-TOF/TOF or nano-ultra-performance liquid chromatography-nanoelectrospray ionization-LTQ-Orbitrap-MS. Direct MALDI-TOF-MS analysis led to the assignment of 57 peptides. Prefractionation by both complementary methods led to the assignment of another 191 peptides. Most peptides originate from α(S1)-casein, followed by β-casein, and α(S2)-casein. κ-Casein and whey proteins seem to play only a minor role as peptide precursors. The formation of many, but not all, peptides could be explained by the activity of the endogenous peptidases, plasmin or cathepsin D, B, and G. Database searches revealed the presence of 22 peptides with established physiological function, including those with angiotensin-converting-enzyme (ACE) inhibitory, immunomodulating, or antimicrobial activity.

The gluten-free, casein-free (GFCF) diet is widely used by families of children with autism spectrum disorders (ASD). Despite its popularity, there is limited evidence in support of the diet. The purpose of this article was to identify and evaluate well-controlled studies of the GFCF diet that have been implemented with children with ASD. A review…

The gluten-free, casein-free (GFCF) diet is widely used by families of children with autism spectrum disorders (ASD). Despite its popularity, there is limited evidence in support of the diet. The purpose of this article was to identify and evaluate well-controlled studies of the GFCF diet that have been implemented with children with ASD. A review…

Many aspects of cellular behavior are affected by information derived from association of the extracellular matrix (ECM) and with cell membrane receptors. When cultured in the presence of laminin-containing ECM and prolactin (Prl), normal mammary epithelial cells express the milk protein beta-casein. Previously, we defined the minimal ECM- and Prl-responsive enhancer element BCE-1 from the upstream region of the beta-casein gene. We also found that BCE-1 was only active when stably integrated into chromatin, and that trichostatin A (TSA), a reagent that leads to alterations in chromatin structure, was able to activate the integrated enhancer element. We now show that endogenous b-casein gene, which is controlled by a genetic assembly that is highly similar to that of BCE-1 and which is also activated by incubation in ECM and Prl, is instead inhibited by TSA. We provide evidence that the differing response of b-casein and BCE-1 to TSA is neither due to an unusual effect of TSA on mammary epithelial cells, nor to secondary consequences from the expression of a separate gene, nor to a particular property of the BCE-1 construct. As a component of this investigation, we also showed that ECM could mediate rapid histone deacetylation in mammary epithelial cells. These results are discussed in combination with previous work showing that TSA mediates the differentiation of many types of cancer cells but inhibits differentiation of some nonmalignant cell types.

protein synthesis (MPS) to intakes of whey and caseinate after heavy resistance exercise in healthy elderly individuals, and, furthermore, to compare the timing effect of caseinate intake. Twenty-four elderly men and women (mean ± SEM; 68 ± 1 years) were randomized to one of four groups: caseinate intake...... before exercise (CasPre), caseinate intake immediately after exercise (CasPost), whey intake immediately after exercise (Whey), or intake of a non-caloric control drink (Control). Muscle myofibrillar and collagen fractional synthesis rates (FSR) were measured by a primed continuous infusion of L-[1......-(13) C]leucine using labeled proteins during a 6-h recovery period. No differences were observed in muscle myofibrillar and collagen FSR with Whey (0.09 ± 0.01%/h) compared with CasPost (0.09 ± 0.003%/h), and it did not differ between CasPre (0.10 ± 0.01%/h) and CasPost. MPS does not differ with whey...

The nucleotide sequence of the cDNA coding for the alpha subunit of casein kinase 2 of Zea mays has been determined. The cDNA clone contains an open reading frame of 996 nucleotides encoding a polypeptide comprising 332 amino acids. The primary amino acid sequence exhibits 75% identity to the alpha...

The proteolytic action of proteases present in cheese plays a major role in the ripening of cheese. These proteases originate from the rennet, the starter cultures and from the milk itself. The proteolysis in cheese results in the degradation of the casein proteins into smaller peptides and free ami

Casein genes have been proved to have an influence on milk properties, and are in addition appropriate for phylogeny studies. A large number of casein polymorphisms exist in goats, making their analysis quite complex. The four casein loci were analyzed by molecular techniques for genetic polymorphism detection in the two dairy goat breeds Bunte Deutsche Edelziege (BDE; n=96), Weisse Deutsche Edelziege (WDE; n=91), and the meat goat breed Buren (n=75). Of the 35 analyzed alleles, 18 were found in BDE, and 17 in Buren goats and WDE. In addition, a new allele was identified at the CSN1S1 locus in the BDE, showing a frequency of 0.05. This variant, named CSN1S1*A', is characterized by a t-->c transversion in intron 9. Linkage disequilibrium was found at the casein haplotype in all three breeds. A total of 30 haplotypes showed frequencies higher than 0.01. In the Buren breed only one haplotype showed a frequency higher than 0.1. The ancestral haplotype B-A-A-B (in the order: CSN1S1-CSN2-CSN1S2-CSN3) occurred in all three breeds, showing a very high frequency (>0.8) in the Buren.

This work reports on the development and characterization of novel meltable polymers and composites based on casein and soybean proteins. The effects of inert (Al2O3) and bioactive (tricalcium phosphate) ceramic reinforcements over the mechanical performance, water absorption, and bioactivity behavi

This work reports on the development and characterization of novel meltable polymers and composites based on casein and soybean proteins. The effects of inert (Al2O3) and bioactive (tricalcium phosphate) ceramic reinforcements over the mechanical performance, water absorption, and bioactivity behavi

Reconstituted skim milks (10 % w/w total solids, pH 6·7-8·0) were ultrasonicated (20, 400 or 1600 kHz at a specific energy input of 286 kJ/kg) at a bulk milk temperature of <30 °C. Application of ultrasound to milk at different pH altered the assembly of the casein micelle in milk, with greater effects at higher pH and lower frequency. Low frequency ultrasound caused greater disruption of casein micelles causing release of protein from the micellar to the serum phase than high frequency. The released protein re-associated to form aggregates of smaller size but with surface charge similar to the casein micelles in the original milk. Ultrasound may be used as a physical intervention to alter the size of the micelles and the partitioning of caseins between the micellar and serum phases in milk. The altered protein equilibria induced by ultrasound treatment may have potential for the development of milk with novel functionality.

To access the fermentative response of equine caecal microbial population to nitrogen availability, an in vitro study was conducted using caecal contents provided with adequate energy sources and nitrogen as limiting nutrient. Two nitrogen (N) sources were provided, protein (casein) and non-protein

Cow's milk allergy is quite common in the first years of human life. Protein composition plays an important role in this pathology, particularly the casein/whey protein ratio. It is known that milks from different species have different sensitization capacities although their protein sources are quite similar. Thus, the objective of this work was to compare the allergenicity of native cow's milk and milk with a modified ratio of casein and whey proteins in a murine model of atopy. Twenty-four Balb/c mice were orally sensitized to native cow's milk or modified cow's milk with a casein/whey protein ratio of 40:60. During the sensitization period, the number of mice suffering from diarrhea was significantly higher in the native cow's milk-sensitized group than in the modified milk-sensitized group. Once mice were killed, plasma histamine levels were shown to be significantly higher in native cow's milk-sensitized mice. In addition, cow's milk proteins induced a higher lymphocyte sensitization in the native milk-sensitized mice, with a significant increase in the specific proliferation ratio of these cells. These results suggest that the balance between caseins and whey proteins plays an important role in the sensitization capacity of cow's milk, and its modification might be a way to reduce the allergenicity of cow's milk.

Full Text Available Objective: Autism is a life long developmental disorder that emerges in early childhood and results in significant lifelong disability. The goal of treatment is to promote the childs social and language development and minimize behaviors that interfere with the childs functioning and learning. This study evaluated the impact of casein and gluten free diet among selected autistic children. Methods: Three private special schools in Salem District, Tamilnadu, India were selected. A total number of 50 autistic children 10 from SIMEC, 10 from MMIC and 30 from CSI comprised the study sample. Background information, clinical history and nutritional status, dietary pattern were collected from the 50 selected autistic children. Out of 50 autistic children 30 autistic children were selected for the dietary intervention. Diet counseling regarding casein free diet was imparted to Group I (n=10, gluten free diet to Group II (n=10 and both casein and gluten free diet for Group III (n=10. The diet was followed for a period of 2 months. The efficacy of the dietary exclusion of casein and gluten was evaluated using a food and behavior diary on a day to day basis, using observation method. Findings: Results about Group I autistic children who followed dietary exclusion of casein free diet showed that the mean scores before and after casein free dietary intervention depiticted these improvements as, 1 to 1.2 for attention, 2.8 to 2.9 for sleep, 1.1 to 1.3 for hyperactivity, 1.1 to 1.2 for anxiety/compulsion. For Group II autistic children who followed dietary exclusion of gluten free diet showed the improvements as 1.1 to 1.4 for attention 2.5 to 3 for sleep, 1.7 to 1.9 for hyperactivity, 1.1 to 1.2 for anxiety/compulsion. About Group III autistic children who followed dietary exclusion of both casein and gluten free diet showed the improvements as 1.1 to 1.3 for attention, 2.5 to 2.7 for sleep, 1.3 to 1.7 for hyperactivity, and 1.1 to 1.2 for anxiety

Full Text Available Objective: to evaluate the efficacy, tolerance, and safety of Carmolis topical gel in patients with gonarthrosis. Subjects and methods. The investigation enrolled 60 patients with knee osteoarthrosis (OA who were divided into two groups: 1 40 patients received Carmolis topical gel in addition to nonsteroidal anti-inflammatory drugs (NSAIDs; 2 20 patients took NSAIDS only (a control group. The treatment duration was 2 weeks. In both groups, therapeutic effectiveness was evaluated from changes in the WOMAC index, pain intensity at rest and during movement by the visual analog scale (VAS. The disease activity was also assessed by a physician and a patient (a Likert scale, local swelling and hyperthermia of the affected joint, the efficiency of treatment, and daily needs for NSAIDs were deter- mined. Results. The performed treatment in both patent groups showed positive clinical changes. Combination therapy involving Carmolis gel displayed greater reductions in WOMAC pain and resting and movement pain than in the con- trol group (as assessed by VAS. On completion of the investigation, considerable improvement was, in the physicians' opinion, noted in 38 (95% patients using Carmolis, which coincided with self-evaluations of the patients. During Carmolis application, the starting dose of NSAIDs could be reduced in 18 (45% patients. Adverse reactions occurred infrequently and required no therapy discontinuation. Conclusion. Carmolis topical gel is effective in relieving clinical symptoms in patients with gonarthrosis, well tolerated, and safe, which can recommend its use in the combination treatment of knee OA.

Milk protein polymorphism plays an important role in genetic diversity analysis, phylogenetic studies, establishing geographical diversity, conservation decision, and improving breeding goals. Milk protein polymorphism in Indian goat breeds has not been well studied; therefore, an investigation was carried out to analyze the genetic structure of the casein gene and milk protein diversity at six milk protein loci in nine Indian goat breeds/genetic groups from varied agro-climatic zones. Milk protein genotyping was carried out in 1098 individual milk samples by SDS-PAGE at alphaS1-CN (CSN1S1), beta-CN (CSN2), alphaS2-CN (CSN1S2), kappa-CN (CSN3), beta-LG, and alpha-LA loci. Indian goats exhibited alphaS1-casein A allele in higher frequency in the majority of breeds except Ganjam and local goats. The alphaS1-casein A allele frequencies varied from 0.45 to 0.77. A total of 16 casein haplotypes were observed in seven breeds and breed specific haplotypes were observed with respect to geographic region. The average number of alleles was lowest in Ganjam (1.66 +/- 0.81) and highest in Sirohi goats (2.50 +/- 1.05). Expected heterozygosity at six different loci demonstrated genetic diversity and breed fragmentation. Neighbor-Joining tree was built basing on Nei's distance. There was about 16.95% variability due to differences between breeds, indicating a strong subdivision. Principal component analysis was carried out to highlight the relationship among breeds. The variability among goat breeds was contributed by alphaS2-CN, beta-LG and alphaS1-CN. The Indian goats exhibited alphaS1-CN (CSN1S1) A allele in higher frequency in all the breeds indicating the higher casein yield in their milk.

Almost all the polyamine-stimulated protein kinase activity associated with the chromatin fraction of nuclei purified from etiolated pea (Pisum sativum L.) plumules is present in a single enzyme that can be extracted from chromatin by 0.35 molar NaCl. This protein kinase can be further purified over 2000-fold by salt fractionation and anion-exchange and casein-agarose column chromatography, after which it is more than 90% pure. The purified kinase has a specific activity of about 650 nanomoles per minute per milligram protein in the absence of polyamines, with either ATP or GTP as phosphoryl donor. Spermidine can stimulate its activity fourfold, with half-maximal activation at about 2 millimolar. Spermine and putrescine also stimulate activity, although somewhat less effectively. This kinase has a tetrameric alpha 2 beta 2 structure with a native molecular weight of 130,000, and subunit molecular weights of 36,000 for the catalytic subunit (alpha) and 29,000 for the regulatory subunit (beta). In western blot analyses, only the alpha subunit reacts strongly with polyclonal antibodies to a Drosophila casein kinase II. The pea kinase can use casein and phosvitin as artificial substrates, phosphorylating both the serine and threonine residues of casein. It has a pH optimum near 8.0, a Vmax of 1.5 micromoles per minute per milligram protein, and a Km for ATP of approximately 75 micromolar. Its activity can be almost completely inhibited by heparin at 5 micrograms per milliliter, but is relatively insensitive to concentrations of staurosporine, K252a, and chlorpromazine that strongly antagonize Ca(2+) -regulated protein kinases. These results are discussed in relation to recent findings that casein kinase 2-type kinases may phosphorylate trans-acting factors that bind to light-regulated promoters in plants.

Full Text Available Abstract Background The mos proto-oncogene encodes a cytoplasmic serine/threonine-specific protein kinase with crucial function during meiotic cell division in vertebrates. Based on oncogenic amino acid substitutions the viral derivative, 124-v-Mos, displays constitutive protein kinase activity and functions independent of unknown upstream effectors of mos protein kinase. We have utilized this property of 124-v-Mos and screened for novel mos substrates in immunocomplex kinase assays in vitro. Results We generated recombinant 124-v-Mos using the baculovirus expression system in Spodoptera frugiperda cells and demonstrated constitutive kinase activity by the ability of 124-v-Mos to auto-phosphorylate and to phosphorylate vimentin, a known substrate of c-Mos. Using this approach we analyzed a panel of acidic and basic substrates in immunocomplex protein kinase assays and identified novel in vitro substrates for 124-v-Mos, the protein tyrosine phosphatase 1B (PTP1B, alpha-casein and beta-casein. We controlled mos-specific phosphorylation of PTP1B and casein in comparative assays using a synthetic kinase-inactive 124-v-Mos mutant and further, tryptic digests of mos-phosphorylated beta-casein identified a phosphopeptide specifically targeted by wild-type 124-v-Mos. Two-dimensional phosphoamino acid analyses showed that 124-v-mos targets serine and threonine residues for phosphorylation in casein at a 1:1 ratio but auto-phosphorylation occurs predominantly on serine residues. Conclusion The mos substrates identified in this study represent a basis to approach the identification of the mos-consensus phosphorylation motif, important for the development of specific inhibitors of the Mos protein kinase.

The objective of this work was to develop and optimize an alternative make process for Greek-style yogurt (GSY), in which the desired level of protein was reached by fortification with micellar casein concentrate (MCC) obtained from milk by microfiltration. Two MCC preparations with 58 and 88% total protein (MCC-58 and MCC-88) were used to fortify yogurt milk to 9.80% (wt/wt) protein. Strained GSY of similar protein content was used as the control. Yogurt milk bases were inoculated with 0.02% (wt/wt) or 0.04% (wt/wt) direct vat set starter culture and fermented until pH 4.5. The acidification rate was faster for the MCC-fortified GSY than for the control, regardless of the inoculation level, which was attributed to the higher nonprotein nitrogen content in the MCC-fortified milk. Steady shear rate rheological analysis indicated a shear-thinning behavior for all GSY samples, which fitted well with the power law model. Dynamic rheological analysis at 5°C showed a weak frequency dependency of the elastic modulus (G') and viscous modulus (G") for all GSY samples, with G' > G", indicating a weak gel structure. Differences in the magnitude of viscoelastic parameters between the 2 types of GSY were found, with G' of MCC-fortified GSY yogurt. Differences were also noticed in water-holding capacity, which was lower for the MCC-fortified GSY compared with the control, attributed to lower serum protein content in the former. Despite some differences in the physicochemical characteristics of the final product compared with GSY manufactured by straining, the alternative process developed here is a feasible alternative to the traditional GSY make process, with environmental and possibly financial benefits to the dairy industry.

Many kinds of stimuli-responsive polymer and gels have been developed and applied to biomimetic actuators or artificial muscles. Electroactive polymers that change shape when stimulated electrically seem to be particularly promising. In all cases, however, the mechanical motion is driven by external stimuli, for example, reversing the direction of electric field. On the other hand, many living organisms can generate an autonomous motion without external driving stimuli like self-beating of he...

The present study examined whether different proteins have different effects on whole-body protein turnover in adult rats. The rats were either starved, given a protein-free but energy-sufficient diet (1 MJ/kg body weight (BW) per d) or a diet containing intact casein, hydrolysed casein...... synthesis. The protein diets produced a positive N balance which was independent of the protein source. Intact and hydrolysed casein increased protein synthesis 2.6- and 2.0-fold respectively, compared with the protein-free diet. Protein degradation increased 1.4- and 1.2-fold respectively. Hydrolysed soya......-bean protein did not increase protein synthesis but decreased protein degradation by 35% compared with the protein-free diet. Compared with the hydrolysed soya-bean protein, intact casein resulted in 2.2- and 2.8-fold higher rates of protein synthesis and degradation respectively. These results are not easily...

This study was aimed at spray drying hydrolysed casein using gum Arabic as the carrier agent, in order to decrease the bitter taste. Three formulations with differing proportions of hydrolysed casein: gum Arabic (10:90, 20:80 and 30:70) were prepared and characterized. They were evaluated for their moisture content, water activity, hygroscopicity, dispersibility in water and in oil, particle size and distribution, particle morphology, thermal behaviour (DSC) and bitter taste by a trained sensory panel using a paired-comparison test (free samples vs. spray dried samples). The proportion of hydrolysed casein did not affect the morphology of the microspheres. The spray drying process increased product stability and modified the dissolution time, but had no effect on the ability of the material to dissolve in either water or oil. The sensory tests showed that the spray drying process using gum Arabic as the carrier was efficient in attenuating or masking the bitter taste of the hydrolysed casein.

Arterial stiffness, blood pressure (BP) and blood lipids may be improved by milk in adults and the effects may be mediated via proteins. However, limited is known about the effects of milk proteins on central aortic BP and no studies have examined the effects in children. Therefore, the present...... stiffness or blood lipid concentrations. A high intake of casein improves DBP in overweight adolescents. Thus, casein may be beneficial for younger overweight subjects in terms of reducing the longterm risk of CVD. In contrast, whey protein seems to increase BP compared with drinking water; however, water...... trial examined the effect of milk and milk proteins on brachial and central aortic BP, blood lipids, inflammation and arterial stiffness in overweight adolescents. A randomised controlled trial was conducted in 193 overweight adolescents aged 12–15 years. They were randomly assigned to drink 1 litre...

An analytical high-performance liquid chromatography (HPLC)-fluorescence method for indirect measuring of whey protein in casein coprecipitate and milk powder was developed. Samples were hydrolyzed with HCl, and cysteyl residues were derivatized with 3,3'-dithiodipropionic acid and 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate. The cysteine content was used to calculate the percentage of whey protein in commercial samples with use of European Union Regulation cysteine reference values in both casein and whey protein. Method validation studies were performed for caseinates and milk powder, and results indicate that the present HPLC approach can be applied as a fast method with a standard deviation of repeatability between 3.3 and 9.5%. Applicability was studied by analysis of 40 commercial caseinate samples, and all complied to European legislation with a content of whey protein not exceeding 5%. Finally, an approach used to estimate the cysteine amount in pure casein by comparison of calculated and experimental values questions the generally accepted cysteine reference value in casein, which is most likely an overestimation.

Full Text Available Polymorphism of goat casein gene closely linked to the quality of milk protein. κ-casein is one of the casein fractions that influence the shape and stability of grain milk. This study is aimed to identify the variation of genotype κ-casein and related with milk quality from Etawah grade, Saanen and their crossbreed (PE-SA. The number of dairy goats used in this study was 150 animals consisted of Etawah grade (48 animals, Saanen (51 animals and PE-SA (51 animals. Steps of experiment were: blood and milk sampling collection, DNA amplification by PCR and the product digestion using Pst1 enzyme, κ-casein gene sequencing and analyzing the quality of fresh milk. The results showed that κ-casein gene is monomorphic by PCR-RFLP (Pst1 for all the goat breeds, but DNA sequencing indicated 38 point of mutation. Observation on goat milk quality showed that Etawah grade milk had highest (P < 0.05 density value (1.033 ± 0.002 and solid non fat (9.577 ± 0.704% than those of Saanen and PE-SA fresh milk goat.

Preterm infants are susceptible to necrotizing enterocolitis (NEC). Using preterm pigs, we determined whether a whey-casein-based formula would be superior to a formula based on whey protein alone. Twenty cesarean-derived preterm pigs (92% gestation) were given total parenteral nutrition for 36 h followed by 30 h of enteral feeding with whey [protein fraction of milk formula based on whey (WHEY); n = 11] or casein and/or whey [protein fraction of milk formula based on a combination of casein and whey (CASEIN); n = 9]-based formulas. Sugar absorptive function was investigated at 6 and 30 h after initiation of enteral feeding using bolus feedings with galactose and mannitol. Pigs were killed after the last in vivo sugar absorption test and evaluated for NEC and the mid intestine was used for ex vivo measurements of hexose absorption. Microbiota profile and short chain fatty acid (SCFA) levels were studied in gut contents. Severity of NEC lesions was similar between diet groups but galactose absorption was markedly higher in CASEIN than in WHEY (P SCFA concentration. Casein may transiently stimulate intestinal sugar absorption but has limited effects on gut structure, microbiota, and NEC in preterm pigs.

In cattle, caseins are subdivided into four main groups: alphaS1-, alphaaS2-, beta- and kappa-caseins. kappa-caseins (CSN2) considerably differ from other caseins in structure and other properties. Testing the A and B alleles is of practical importance, because the milk of cows that carry the B allele of CSN3 has a better thermal resistance and shorter coagulation time, better curdles and contains micelles of different sizes. Iranian Sistani cattle (Bos indicus) are a heavy built breed and used as dual-purpose cattle breed in Eastern Iran. This breed is a genetic resource that shows special features of adaptation to rustic environments. One of the most distinctive features of Sistani cattle is its great capability to resist diseases which makes it a potential reservoir of germplasm useful for future crosses. Our main goal was to study DNA-polymorphism of the CSN3 gene in the Iranian Sistani native cattle (Bos indicus) and analyze the information value of CSN3 gene polymorphism as a genetic marker. We genotyped and analyzed 65 animals of this breed using PCR-RFLP. The frequencies of A and B alleles were 0.6385 and 0.3615 and those of AA, AB and BB genotypes were 0.4000, 0.4769 and 0.1231, respectively. In the Sistani Zebu breed, frequency of B allele is higher than other Zebu breeds, indicating that intensive selection for dairy production have been done and indirectly influenced CSN3 allele frequencies.

The main goal of the current work was to identify single nucleotide polymorphisms (SNP) that might create or disrupt microRNA (miRNA) target sites in the caprine casein genes. The 3' untranslated regions of the goat alpha(S1)-, alpha(S2)-, beta-, and kappa-casein genes (CSN1S1, CSN1S2, CSN2, and CSN3, respectively) were resequenced in 25 individuals of the Murciano-Granadina, Cashmere, Canarian, Saanen, and Sahelian breeds. Five SNP were identified through this strategy: c.175C>T at CSN1S1; c.109T>C, c.139G>C, and c.160T>C at CSN1S2; and c.216C>T at CSN2. Analysis with the Patrocles Finder tool predicted that all of these SNP are located within regions complementary to the seed of diverse miRNA sequences. These in silico results suggest that polymorphism at miRNA target sites might have some effect on casein expression. We explored this issue by genotyping the c.175C>T SNP (CSN1S1) in 85 Murciano-Granadina goats with records for milk CSN1S1 concentrations. This substitution destroys a putative target site for miR-101, a miRNA known to be expressed in the bovine mammary gland. Although TT goats had higher levels (6.25 g/L) of CSN1S1 than their CT (6.05 g/L) and CC (6.04 g/L) counterparts, these differences were not significant. Experimental confirmation of the miRNA target sites predicted in the current work and performance of additional association analyses in other goat populations will be an essential step to find out if polymorphic miRNA target sites constitute an important source of variation in casein expression.

Foam stability of casein micelle dispersions (CMDs) strongly depends on aggregate size. To elucidate the underlying mechanism, the role of interfacial and thin film properties was investigated. CMDs were prepared at 4°C and 20°C, designated as CMD4°C and CMD20°C. At equal protein concentrations, foam stability of CMD4°C (with casein micelle aggregates) was markedly higher than CMD20°C (without aggregates). Although the elastic modulus of CMD4°C was twice as that of CMD20°C at 0.005Hz, the protein adsorbed amount was slightly higher for CMD20°C than for CMD4°C, which indicated a slight difference in interfacial composition of the air/water interface. Non-linear surface dilatational rheology showed minor differences between mechanical properties of air/water interfaces stabilized by two CMDs. These differences in interfacial properties could not explain the large difference in foam stability between two CMDs. Thin film analysis showed that films made with CMD20°C drained to a more homogeneous film compared to films stabilized by CMD4°C. Large casein micelle aggregates trapped in the thin film of CMD4°C made the film more heterogeneous. The rupture time of thin films was significantly longer for CMD4°C (>1h) than for CMD20°C (foam stability decreased significantly. In conclusion, the increased stability of foam prepared with CMD4°C appears to be the result of entrapment of casein micelle aggregates in the liquid films of the foam.

There are inconsistent reports on the lactose and/or galactose content of some foods traditionally restricted from the diet for classic galactosemia. Therefore, samples of cheeses, caseinates, and canned black, pinto, kidney, and garbanzo beans were analyzed for free galactose content using HPLC with refractive index or pulsed amperometric detection. Galactose concentrations in several hard and aged cheeses and three mild/medium Cheddars, produced by smaller local dairies, was galactosemia.

By combining the osmotic stress technique with small-angle x-ray scattering measurements, we followed the structural response of the casein micelle to an overall increase in concentration. When the aqueous phase that separates the micelles is extracted, they behave as polydisperse repelling spheres and their internal structure is not affected. When they are compressed, the micelles lose water and shrink to a smaller volume. Our results indicate that this compression is nonaffine, i.e., some p...

Background: Autism is a complex psychiatric disorder characterized by three core symptoms, i.e. impairments in social interaction, restricted patterns of behavior and impairments in communication. In the framework of the “opioid excess theory”, the disorder symptoms are compared to the behavioral effects of opiates. Based on this, a possible nutritional basis of autism has been proposed, hypothesizing that certain food proteins such as gluten and casein can be transformed ...

Alginate remains the most popular polymer used for cell encapsulation, yet its biocompatibility is inconsistent. Two commercially available alginates were compared, one with 71% guluronate (HiG), and the other with 44% (IntG). Both alginates were purified, and their purities were verified. After 2 days in the peritoneal cavity of C57BL/6J mice, barium (Ba)-gel and calcium (Ca)-gel beads of IntG alginate were clean, while host cells were adhered to beads of HiG alginate. IntG gel beads, however, showed fragmentation in vivo while HiG gel beads stayed firm. The physicochemical properties of the sodium alginates and their gels were thoroughly characterized. The intrinsic viscosity of IntG alginate was 2.5-fold higher than that of HiG alginate, suggesting a greater molecular mass. X-ray photoelectron spectroscopy indicated that both alginates were similar in elemental composition, including low levels of counterions in all gels. The wettabilities of the alginates and gels were also identical, as measured by contact angles of water on dry films. Ba-gel beads of HiG alginate resisted swelling and degradation when immersed in water, much more than the other gel beads. These results suggest that the main factors contributing to the biocompatibility of gels of purified alginate are the mannuronate/guluronate content and/or intrinsic viscosity.

Skim milk was heated at .70, 95, and 140{degree}C to simulate the processes of pasteurization, forewarming, and UHT sterilization, and the specific interactions between {alpha}-lactalbumin or {beta}-lactoglobulin and the caseins studied using tracer amounts of added {sup 14}C-labeled whey protein. Radioactivities of the whey and of the washed casein pellets from renneted skim milk were measured and the extent of the interaction estimated. Upon heating skim milk at 70{degree}C for 45 s, less than 2% {beta}-lactoglobulin and less than .3% {alpha}-lactalbumin were incorporated into the curd. Heating at 95{degree}C for .5 to 20 min resulted in 58 to 85% of the {beta}-lactoglobulin and 8 to 55% of the {alpha}-lactalbumin becoming associated with the curd. Heating at 140{degree}C for 2 and 4 s caused 43 and 54% of the {beta}-lactoglobulin and 9 and 12% of the {alpha}-lactalbumin, respectively, to be bound to the curd fraction. The radiolabeling technique is very sensitive and useful for tracing low levels of interaction between whey proteins and casein in heated milk systems.

"Requeijão cremoso" is a traditional Brazilian processed cheese spread, showing ample acceptance on the national market. Emulsifying salts (ES) are an important factor influencing the characteristics of processed cheeses, but the literature presents conflicting results about their action on cheese functionality. Requeijão cremoso obtained from anhydrous ingredients allows the study of the influence of each type of ES on the cheese properties, since it can be treated as a model system where the variables are limited and well known. The objective of this study was to evaluate the effect of different types of ES (TSC-sodium citrate, SHMP-sodium hexametaphosphate, STPP-sodium tripolyphosphate, and TSPP-tetrasodium pyrophosphate) on the sensory and functional characteristics of requeijão cremoso-processed cheeses obtained from anhydrous ingredients. The physicochemical composition, degree of casein dissociation, fat particle size, melting index, color, texture profile, and sensory acceptance of the cheeses were determined. The functional behavior of processed cheeses was strongly influenced by the type of ES and its physicochemical properties including its ability to bind Ca, the casein dispersion during cooking, and the possible creation of cross-links with casein during cooling. The cheese made with SHMP was the one most differentiated from the others, presenting lower melting index, whiter color, and higher values for hardness, gumminess, and adhesiveness. The differences in texture had an impact on sensory acceptance: with the exception of the sample manufactured with sodium hexametaphosphate, all the samples presented good sensory acceptance.

This study investigated the formation of N(ε)-carboxymethyllysine (CML) in two caseinate solutions containing: (1) glucose, (2) lactose, each heated at 120 °C and 130 °C. At both heating temperatures, CML concentration in lactose-caseinate solution was higher than in glucose-caseinate solution. In both solutions, more CML was formed at 130 °C than at 120 °C. Using multiresponse modelling, two degradation routes for the sugars were confirmed: (1) isomerisation of glucose or lactose and subsequent degradation via Lobry de Bruyn-Alberda van Ekenstein (LA) arrangement; (2) the Maillard reaction between the reducing sugar and lysine residues. Modelling results suggested that CML was not formed from oxidation of the reducing sugars, but from the Maillard reaction via the Amadori rearrangement product. Since CML appeared to be thermally unstable under the current study conditions, it may not be a perfect indicator for heat damage of processed foods. This is the first study in which CML formation was linked to available information on the Maillard reaction via multiresponse modelling.

Full Text Available Aim: Yak milk is a type of milk that people are less familiar with due to its remote geographical location which may havesignificant effects on composition, microbiota and hydrolytic outcome. Present work was designed with the aim to evaluatethe antioxidative effect of peptides derived from yak milk caseinate on hydrolysis with three different proteases.Materials and Methods: In this investigation Yak milk casein was hydrolyzed by three commercially available proteases(Trypsin, Pepsin and chymotrypsin. These hydrolysates collected at different hydrolysis times (30 min, 60 min, 90 min, 120min, 150 min, 180 min, 210 min, 240 min, 270 min, 300 min, 330 min and 360 min were assayed for their antioxidant activitywith respect to the effect of incubation period.Results: Among all the enzyme hydrolysates, the tryptic hydrolysates showed highest antioxidant activity followed bychymotryptic hydrolysates. Further, the peptide samples showing highest activity were subjected to RP-HPLC for their partialcharacterization. Tryptic and peptic hydrolysates produced peaks mainly in the region of hydrophillic solvent indicating thepresence of hydrophillic peptides/peptides.Conclusion: The results indicated that yak milk casein could be a resource to generate antioxidative peptides and be used asmultifunctional active ingredients for many value-added functional foods as well as a traditional food protein.

Full Text Available A total of 137,753 test day records of 20,745 Italian Brown Swiss dairy cows from 26 provinces of Italy were used to estimate heritability for casein and urea content in milk and their genetic correlations with other production traits and milk somatic cell score. Milk component values were obtained by Fourier Transformed Infrared (IR Spectroscopy from milk samples collected during national routine recording and were analysed using test day repeatability animal models. Fixed effects included 1,001 levels of herd-test date, 15 classes of days in milk, and 13 classes of age at calving within parity. The variation among cows was large for most of the traits. The heritability value for casein content was 0.31, as for protein content, and genetic and phenotypic correlations between these two traits were large (0.99 and 0.97 respectively. Milk urea content had a heritability of 0.17 and a positive genetic relationship with fat (0.12, null with protein (0.03 and casein (0.002 content and a negative genetic correlation with milk yield (-0.17 suggesting that the genetic improvement for milk urea content would be possible, but genetic gain would be affected by other traits included as selection criteria in the economic index and by their relative economic emphasis.

In this work, a novel encapsulation method was studied by spray-drying a warm aqueous ethanol solution with codissolved sodium caseinate (NaCas) and lipophilic food components, using curcumin as a model compound. The encapsulation caused the loss of crystallinity of curcumin. After hydration of spray-dried powder and centrifugation, 137 μg/mL curcumin was dispersed in the transparent dispersion, which was 4 decades higher than its water solubility. Dynamic light scattering and atomic force microscopy results showed that curcumin-loaded casein nanoparticles were bigger than those of NaCas processed at encapsulation conditions but were smaller than those of the native NaCas. The increased nanoparticle dimension, together with fluorescence and FTIR spectroscopy results, suggested that curcumin was entrapped in the nanoparticle core through hydrophobic interactions. The curcumin encapsulated in casein nanoparticles had higher biological activity, as assessed by antioxidant and cell proliferation assays, than pristine curcumin, likely due to the improved dispersibility. This simple approach may be applied to encapsulate various lipophilic bioactive compounds.

Biosurfactants are proteins with detergent, emulsifier, and antimicrobial actions that have potential application in environmental applications such as the treatment of organic pollutants and oil recovery. Bacillus atrophaeus strains are nonpathogenic and are suitable source of biosurfactants, among which is surfactin. The aim of this work is to establish a culture medium composition able to stimulate biosurfactants production by B. atrophaeus ATCC 9372. Batch cultivations were carried out in a rotary shaker at 150 rpm and 35°C for 24 h on glucose- and/or casein-based semidefined culture media also containing sodium chloride, dibasic sodium phosphate, and soy flour. The addition of 14.0 g/L glucose in a culture medium containing 10.0 g/L of casein resulted in 17 times higher biosurfactant production (B max=635.0 mg/L). Besides, the simultaneous presence of digested casein (10.0 g/L), digested soy flour (3.0 g/L), and glucose (18.0 g/L) in the medium was responsible for a diauxic effect during cell growth. Once the diauxie started, the average biosurfactants concentration was 16.8% less than that observed before this phenomenon. The capability of B. atrophaeus strain to adapt its own metabolism to use several nutrients as energy sources and to preserve high levels of biosurfactants in the medium during the stationary phase is a promising feature for its possible application in biological treatments.

Full Text Available Dietary intervention as a tool for maintaining and improving physical health and wellbeing is a widely researched and discussed topic. Speculation that diet may similarly affect mental health and wellbeing particularly in cases of psychiatric and behavioural symptomatology opens up various avenues for potentially improving quality of life. We examine evidence suggestive that a gluten-free, casein-free or combined gluten- and casein-free diet can ameliorate core and peripheral symptoms and improve developmental outcome in some cases of autism spectrum conditions. Although not wholly affirmative, the majority of published studies indicate statistically significant positive changes to symptom presentation following dietary intervention. In particular, changes to areas of communication, attention and hyperactivity are detailed, despite the presence of various methodological shortcomings. Specific characteristics of best- and non-responders to intervention have not been fully elucidated; neither has the precise mode of action for any universal effect outside of known individual cases of food-related co-morbidity. With the publication of controlled medium- and long-term group studies of a gluten- and casein-free diet alongside more consolidated biological findings potentially linked to intervention, the appearance of a possible diet-related autism phenotype seems to be emerging supportive of a positive dietary effect in some cases. Further debate on whether such dietary intervention should form part of best practice guidelines for autism spectrum conditions and onward representative of an autism dietary-sensitive enteropathy is warranted.

Full Text Available The article presents the results of studies on the functional properties of carrageenan, depending on the concentration of sodium chloride and xanthan in gels. It is established that the main factors in the syneresis of carrageenan gels are its concentration, the presence of ions and gums in solution. If using sodium chloride there is a change in the structure of mesh of the resulting gel, which leads to an increase in syneresis.

Nanostructured gel polymer electrolytes that have both high ionic conductivity and high mechanical strength are disclosed. The electrolytes have at least two domains--one domain contains an ionically-conductive gel polymer and the other domain contains a rigid polymer that provides structure for the electrolyte. The domains are formed by block copolymers. The first block provides a polymer matrix that may or may not be conductive on by itself, but that can soak up a liquid electrolyte, thereby making a gel. An exemplary nanostructured gel polymer electrolyte has an ionic conductivity of at least 1.times.10.sup.-4 S cm.sup.-1 at 25.degree. C.

Caseins,the main components of milk protein, represent about 80% of the total protein in the milk.Owing to the importance of casein and casein micelles for the functional behavior of dairy products, and widely application of caseins, the study of casein is always drawing more attention.Especially, the exact structure of casein micelles is still under debate.Many scholars have proposed various theoretic models for casein micelle structure. This paper reviewed kinds means of casein micelles structure,dissociation and aggregation in order to provide a reference for studying casein in the future.%酪蛋白是乳中蛋白质的主要成分，约占其80％。由于酪蛋白胶束对乳制品的功能特性起重要作用，因此其研究一直备受关注，但酪蛋白胶束的内部结构至今没有确切的定义，而是许多学者提出了各种理论模型。文章综述了酪蛋白胶束结构、解离与聚集的多种研究方法，为以后学者研究酪蛋白提供借鉴。

The aim of this study was to introduce a reference method for the detection of cow milk in ewe and goat cheeses (EC 273/08) in order to protect manufacturers and consumers from adulterations and imitations, and to ensure alignment with the demands of domestic and EU markets. The method includes isolation of casein from cheese, isoelectric focusing of γ2- and γ3-casein originating from the hydrolysis of β-casein by plasmin, the detection and quantitative determination of γ-casein in cow, ewe a...

A novel water-soluble 1,8-naphthalimide derivative 1, bearing two acetic carboxylic groups, exhibited fluorescent turn-on recognition for casein micelle based on the aggregation induced emission (AIE) character. The photophysical properties of 1 consisting of donor and acceptor units were investigated by the solvation effect. Changing from polar to non-polar solvent increased the solvent interaction; both the excitation and emission spectra were shifted to shorter wavelength and intensity decreased through taking advantage of twisted intramolecular charge transfer (TICT) and self-association fluorescence emission. Moreover, the red-shift and quenching in protic solvent were caused by the excited-state hydrogen bond strengthening effect. The density functional theory (DFT) and time dependent density functional theory (TDDFT) were used to obtain the most stable structure, electronic excitation energy, dipole moments and charge distribution. The AIE mechanism of 1 with casein micelle was due to 1 docked in the hydrophobic cavity between sub-micelles and bound with amino acid residues, resulting in the aggregation of 1 on the casein micelle surface and emission enhancement, based on which, a novel casein assay method was developed. The proposed method exhibited a good linear range from 0.1 to 10.5 μg mL{sup −1}, with the detection limit of 3.0 ng mL{sup −1}. Satisfactory reproducibility, reversibility and a short response time were realized. This method was applied for the determination of casein in milk powder samples, avoiding the interferences from other components and illegal additives in milk. -- Highlights: • A water-soluble 1,8-naphthalimide-based fluorescent probe 1 was synthesized. • Photophysical characterization of 1 was studied. • Aggregation induced emission enhancement of 1 with casein was investigated. • A novel casein quantification method was developed.

Mammary alveolar (MAC-T) cells, an established bovine mammary epithelial cell line, are frequently used to investigate differentiation. A lactogenic phenotype in these cells is induced by treatment with a combination of hydrocortisone, insulin, and prolactin (PRL). The effect of the vitamin A derivative retinoic acid (RA), which induces differentiation in many cells, has not been studied in MAC-T cells. The objective of this study was to evaluate the differentiation potential of RA (1 μM) in MAC-T cells and to examine the effect of combined treatment with RA (1 μM) and PRL (5 μg/mL). Although RA treatment alone inhibited MAC-T cell proliferation, co-treatment of RA with PRL increased cell growth compared with the control group (treated with 1 μg/mL hydrocortisone and 5 μg/mL insulin). The ratio of Bcl to Bax mRNA was decreased in the RA treatment compared with RA+PRL or control. Retinoic acid-induced differentiation of MAC-T cells was associated with an increase in the mRNA expression of αS1-casein (3.9-fold), αS2-casein (4.5-fold), and β-casein (4.4-fold) compared with the control group. Expression of αS1-casein, αS2-casein, and β-casein was increased 12.9-fold, 11.9-fold, and 19.3-fold, respectively, following treatment with RA and PRL combined compared with the control group. These results demonstrate that RA induces differentiation of MAC-T cells and acts synergistically with PRL to increase specific casein gene expression.

Full Text Available The production and consumption of mare's milk in Europe has gained importance, mainly based on positive health effects and a lower allergenic potential as compared to cows' milk. The allergenicity of milk is to a certain extent affected by different genetic variants. In classical dairy species, much research has been conducted into the genetic variability of milk proteins, but the knowledge in horses is scarce. Here, we characterize two major forms of equine αS2-casein arising from genomic 1.3 kb in-frame deletion involving two coding exons, one of which represents an equid specific duplication. Findings at the DNA-level have been verified by cDNA sequencing from horse milk of mares with different genotypes. At the protein-level, we were able to show by SDS-page and in-gel digestion with subsequent LC-MS analysis that both proteins are actually expressed. The comparison with published sequences of other equids revealed that the deletion has probably occurred before the ancestor of present-day asses and zebras diverged from the horse lineage.

Full Text Available Molar Incisor Hypomineralization (MIH is characterized by a developmentally derived deficiency in mineral enamel. Affected teeth present demarcated enamel opacities, ranging from white to brown; also hypoplasia can be associated. Patient frequently claims aesthetic discomfort if anterior teeth are involved. This problem leads patients to request a bleaching treatment to improve aestheticconditions.Nevertheless, hydrogen peroxide can produce serious side-effects, resulting from further mineral loss. Microabrasion and/or a composite restoration are the treatments of choice in teeth with mild/moderate MIH, but they also need enamel loss. Recently, a new remineralizing agent based on Casein Phosphopeptide-Amorphous Calcium Phosphate (CPP-ACP has been proposed to be effective in hypomineralized enamel, improving also aesthetic conditions. The present paper presents a case report of a young man with white opacities on incisors treated with a combined use of CPP-ACP mousse and hydrogen peroxide gel to correct the aesthetic defect. The patient was instructed to use CPP-ACP for two hours per day for three months in order to obtain enamel remineralization followed by a combined use of CPP-ACP and bleaching agent for further two months. At the end of this five-month treatment, a noticeable aesthetic improvement of the opacities was observed.

Molar Incisor Hypomineralization (MIH) is characterized by a developmentally derived deficiency in mineral enamel. Affected teeth present demarcated enamel opacities, ranging from white to brown; also hypoplasia can be associated. Patient frequently claims aesthetic discomfort if anterior teeth are involved. This problem leads patients to request a bleaching treatment to improve aestheticconditions.Nevertheless, hydrogen peroxide can produce serious side-effects, resulting from further mineral loss. Microabrasion and/or a composite restoration are the treatments of choice in teeth with mild/moderate MIH, but they also need enamel loss. Recently, a new remineralizing agent based on Casein Phosphopeptide-Amorphous Calcium Phosphate (CPP-ACP) has been proposed to be effective in hypomineralized enamel, improving also aesthetic conditions. The present paper presents a case report of a young man with white opacities on incisors treated with a combined use of CPP-ACP mousse and hydrogen peroxide gel to correct the aesthetic defect. The patient was instructed to use CPP-ACP for two hours per day for three months in order to obtain enamel remineralization followed by a combined use of CPP-ACP and bleaching agent for further two months. At the end of this five-month treatment, a noticeable aesthetic improvement of the opacities was observed.

Gelation kinetics, mechanical spectra, thermal scanning rheology (TSR), and differential scanning calorimetry (DSC) in aqueous solutions of gelling polymers and colloids such as seaweed polysaccharides (agarose, carrageenans), microbial polysaccharides (gellan, curdlan), plant polysaccharides (methylcellulose), globular proteins (casein, glycinin, {beta}-conglycinin), fibrous proteins (gelatin, fibrin), and polyvinyl alcohol, which are related to foods, cosmetics, biomedical and pharmaceutical applications, are described. Some gelation processes at a constant temperature have been treated successfully by an equation of first order kinetics or by other modified equations, and the molecular mechanism of gel formation is discussed briefly. For water-soluble polymers, the criterion of the gel or sol based on the frequency dependence of storage and loss moduli gives valuable informations. TSR and DSC are complementary, and the combination of these methods has been proved to be useful. (orig.) 81 refs.

The present invention is directed to solid composites including colloidal nanocrystals within a sol-gel host or matrix and to processes of forming such solid composites. The present invention is further directed to alcohol soluble colloidal nanocrystals useful in formation of sol-gel based solid composites.

The equine alpha(s1)- and beta-caseins (CN) were purified by chromatography on DEAE-cellulose and by reversed-phase HPLC. The alpha(s1)-, beta-, and kappa-CN were characterized either by monodimensional urea-PAGE or sodium dodecylsulfate (SDS)-PAGE or by bidimensional electrophoresis. Kappa-casein was characterized after electrophoresis by glycoprotein-specific staining. To identify alpha(s1)-CN without ambiguity, internal sequences were determined after trypsin or chymosin digestion of purified alpha(s1)-CN. These sequences, that could be estimated to correspond to 62% of the full protein, presented strong identities with regions of alpha(s1)-CN primary structures of other species. In particular, 51, 48, 43, and 40% identities were obtained with corresponding regions of sow, dromedary, cow, and human alpha(s1)-CN, respectively. On the other hand, trace amounts of equine gamma-CN-like and proteose peptone component 5-like peptides were found in the whole CN. They were identified by microsequencing and corresponded to beta-CN peptides generated by plasmin action on the whole CN. The equine alpha(s1), beta-, and kappa-CN were separated by bidimensional electrophoresis in numerous isoelectric variants with apparent isoelectric points distributed between pH 4.4 to 6.3, 4.4 to 5.9, and 3.5 to 5.5, respectively. The beta- and kappa-CN displayed a more acidic character in the mare than in the cow.

Sucrose release from polysaccharide gels has been studied extensively because it is expected to be useful in understanding flavour release from solid foods and to find a new processing method which produces more palatable and healthier foods. We provide an overview of the release of sucrose and other sugars from gels of agar and related polysaccharides. The addition of sucrose to agar solutions leads to the increase in transparency of the resulting gels and the decrease in syneresis, which is attributed to the decrease in mesh size in gels. The syneresis occurring in the quiescent condition and fluid release induced by compression is discussed. The relationship between the sugar release and the structural, rheological and thermal properties of gels is also discussed. Finally, the future research direction is proposed.

Full Text Available Light-emitting gel, a gel state electroluminescence material, is reported. It is composed of a ruthenium complex as the emitter, an ionic liquid as the electrolyte, and oxide nanoparticles as the gelation filler. Emitted light was produced via electrogenerated chemiluminescence. The light-emitting gel operated at low voltage when an alternating current was passed through it, regardless of its structure, which is quite thick. The luminescence property of the gel is strongly affected by nanoparticle materials. TiO2 nanoparticles were a better gelation filler than silica or ZnO was, with respect to luminescence stability, thus indicating a catalytic effect. It is demonstrated that the light-emitting gel device, with quite a simple fabrication process, flashes with the application of voltage.

Functional properties of heat-induced gels prepared from microbial transglutaminase (TG)-treated porcine myofibrillar protein (MP) containing sodium caseinate with or without konjac flour (KF) under various salt concentrations (0.1, 0.3 and 0.6MNaCl) were evaluated. The mixed MP gels with KF exhibited improved cooking yields at all salt concentrations. TG treatment greatly enhanced gel strength and elasticity (storage modulus, G') at 0.6M NaCl, but not at lower salt concentrations. The combination of KF and TG improved the gel strength at 0.1 and 0.3M NaCl and G' at all salt concentrations, when compared with non-TG controls. Incubation of MP suspensions (sols) with TG promoted the disappearance of myosin heavy chain and the production of polymers. The TG-treated MP mixed gels had a compact structure, compared to those without TG, and the KF incorporation modified the gel matrix and increased its water-holding capacity. Results from differential scanning calorimetry suggested possible interactions of MP with KF, which may explain the changes in the microstructure of the heat-induced gels.

The effect of tetrasodium pyrophosphate (TSPP) on the properties of yogurt gels was investigated. Various concentrations (0.05 to 0.2%) of TSPP were added to preheated (85 degrees C for 30 min) reconstituted skim milk, which was readjusted to pH 6.50. Milk was inoculated with 2% starter culture and incubated at 42 degrees C until the pH reached 4.6. Acid-base buffering profiles of milk and total and soluble calcium levels were measured. Turbidity measurements were used to indicate changes in casein dispersion. Storage modulus (G') and loss tangent (LT) values of yogurts were monitored during fermentation using dynamic oscillatory rheology. Large deformation properties of gels were also measured. Microstructural properties of yogurt were observed using fluorescence microscopy. The addition of TSPP resulted in the disappearance of the buffering peak during acid titration at pH approximately 5.1 that is due to the solubilization of colloidal calcium phosphate (CCP), and a new peak was observed at lower pH values (pH 4.0-4.5). The buffering peak at pH 6.0 during base titration virtually disappeared with addition of TSPP and a new peak appeared at pH approximately 4.8. The addition of TSPP reduced the soluble Ca content of milk and increased casein-bound Ca values. The addition of up to 0.125% TSPP resulted in a reduction in turbidity because of micelle dispersion but at 0.15%, turbidity increased and these samples exhibited a time-dependent increase in turbidity because of aggregation of casein particles. Gels made with 0.20% TSPP were very weak and had a very high gelation pH (6.35), probably due to complete dispersion of the micelle structure in this sample. The LT value of gels at pH 5.1 decreased with an increase in TSPP concentration, probably due to the loss of CCP with the addition of TSPP. The G' values at pH 4.6 of gels made with or=0.125% TSPP significantly decreased G' values. The addition of 0.05 to 0.125% TSPP to milk resulted in a reduction in the yield

The relationship between genetic variants for milk protein and the composition of milk was analyzed on 4475 repeated milk samples from individual cows; 371 dairy cows of the Swedish Red and White breed and 204 cows of the Swedish Holstein breed were used. The registrations included percentages of casein, protein, fat, and lactose in combination with milk yield and SCC. The genotype of individual cows for alpha(s1)-CN, beta-CN, kappa-CN, and beta-LG was determined by alkaline and acidic PAGE. A mixed animal model was used for the analysis; beta-LG and aggregate casein genotypes were included simultaneously as separate fixed effects in the statistical model. The results suggest a positive additive effect of the beta-LG B allele on casein content and on the ratio of casein to total protein. For the latter trait, the beta-LG genotype accounted for a relatively large part of the phenotypic variance, corresponding to a reduction in residual variance of 11% when included in the model. The corresponding value for casein content was 0.5%. The lack of unfavorable associations between milk protein variants and the traits included in this study makes the beta-LG gene an obvious candidate when the breeding objective is improved conversion of milk protein into cheese.

Full Text Available Abstract κ-casein plays an essential role in the formation, stabilisation and aggregation of milk micelles. Control of κ-casein expression reflects this essential role, although an understanding of the mechanisms involved lags behind that of the other milk protein genes. We determined the 5'-flanking sequences for the murine, rabbit and human κ-casein genes and compared them to the published ruminant sequences. The most conserved region was not the proximal promoter region but an approximately 400 bp long region centred 800 bp upstream of the TATA box. This region contained two highly conserved MGF/STAT5 sites with common spacing relative to each other. In this region, six conserved short stretches of similarity were also found which did not correspond to known transcription factor consensus sites. On the contrary to ruminant and human 5' regulatory sequences, the rabbit and murine 5'-flanking regions did not harbour any kind of repetitive elements. We generated a phylogenetic tree of the six species based on multiple alignment of the κ-casein sequences. This study identified conserved candidate transcriptional regulatory elements within the κ-casein gene promoter.

In this work, two proteins, Z-domains and bovine casein, were auto-displayed on the outer membrane of the same Escherichia coli cells by co-transformation of two different auto-display vectors. On the basis of SDS-PAGE densitometry, Z-domains and bovine casein were expressed at 3.12 × 10⁵ and 1.55 × 10⁵ proteins/E. coli cell, respectively. The co-auto-displayed Z-domains had antibody-binding activity and the bovine casein had adhesive properties. E. coli with co-auto-displayed proteins were analyzed by fluorescence assisted cell sorting (FACS). E. coli with co-auto-displayed Z-domains and bovine casein aggregated due to hydrophobic interaction. For application to immunoassays, the Z-domain activity was estimated after (1) immobilizing the E. coli and (2) forming an OM layer. E. coli with co-auto-displayed two proteins that were immobilized on a polystyrene microplate had the same antibody-binding activity as did E. coli with auto-displayed Z-domains only. The OM layer from the co-transformed E. coli had Z-domains and bovine casein expressed at a 1:2 ratio from antibody-binding activity measurements.

The interaction of bis(indolyl)methane with bovine milk β-casein was investigated using spectroscopy and molecular docking studies at different temperatures (25-37 °C). The circular dichroism and Fourier transform infrared spectroscopic data demonstrated that β-casein interacts with BIM molecule mainly via both the hydrophobic and hydrophilic interactions with a minor change in the secondary structure of β-casein. The fluorescence quenching measurements revealed that the presence of a single binding site on β-casein for BIM with the binding constant value of ∼104 M-1. The negative values of entropy and enthalpy changes confirm the predominate role of hydrogen binding and van der Waals interactions in the binding process. Fӧrster energy transfer measurement suggested that the distance between bound BIM and Trp residue is higher than the respective critical distance. Hence, the static quenching is more likely responsible for the fluorescence quenching rather than the mechanism of non-radiative. Docking study showed that BIM molecule forms three hydrogen bonds and several van der Waals contacts with β-casein.

The recoverable energy (RE), defined as the ratio of the work exerted on a test specimen during compression and recovered upon subsequent decompression, has been shown to correlate to sensory profiling of protein-based food products. Understanding the mechanism determining the time-dependency of RE is primordial. This work aims to identify the protein-specific impact on the recoverable energy by stress dissipation via relaxation of (micro)structural rearrangements within protein gels. To this end, caseinate and gelatin gels are studied for their response to time-dependent mechanical deformation as they are known to develop structurally distinct network morphologies. This work shows that in gelatin gels no significant stress relaxation occurs on the seconds timescale, and consequently no time-dependency of the amount of energy stored in this material is observed. In caseinategels, however, the energy dissipation via relaxation processes does contribute significantly to the time-dependency of reversible stored energy in the network. This can explain the obtained RE as a function of applied deformation at slow deformation rates. At faster deformation, an additional contribution to the dissipated energy is apparent, that increases with the deformation rate, which might point to the role of energy dissipation related to friction of the serum entrapped by the protein-network. This work shows that engineering strategies focused on controlling viscous flow in protein gels could be more effective to dictate the ability to elastically store energy in protein gels than routes that direct protein-specific aggregation and/or network-assembly.

A high resolution SNP map was constructed for the bovine casein region to identify haplotype structures and study associations with milk traits in Norwegian Red cattle. Our analyses suggest separation of the casein cluster into two haplotype blocks, one consisting of the CSN1S1, CSN2 and CSN1S2 genes and another one consisting of the CSN3 gene. Highly significant associations with both protein and milk yield were found for both single SNPs and haplotypes within the CSN1S1-CSN2-CSN1S2 haplotype block. In contrast, no significant association was found for single SNPs or haplotypes within the CSN3 block. Our results point towards CSN2 and CSN1S2 as the most likely loci harbouring the underlying causative DNA variation. In our study, the most significant results were found for the SNP CSN2_67 with the C allele consistently associated with both higher protein and milk yields. CSN2_67 calls a C to an A substitution at codon 67 in beta-casein gene resulting in histidine replacing proline in the amino acid sequence. This polymorphism determines the protein variants A1/B (CSN2_67 A allele) versus A2/A3 (CSN2_67 C allele). Other studies have suggested that a high consumption of A1/B milk may affect human health by increasing the risk of diabetes and heart diseases. Altogether these results argue for an increase in the frequency of the CSN2_67 C allele or haplotypes containing this allele in the Norwegian Red cattle population by selective breeding.

Full Text Available Kappa casein (ĸ-cn gene as one of the four casein genes commonly has two variant alleles. B allele is positively correlated with milk proteins, so potentially be used as MAS to improve milk protein content. This study aimed to identify genetic variation of the ĸ-casein gene in Holstein-Friesian (HF from several dairy regions in West Java. Blood samples were taken through jugular vein of lactating heifers and cows, from intensive managements under the Local Government Dairy Breeding Station - Cikole (BBPT Cikole-SP (82 hds, Embryo Transfer Station (BET (50 hds, and Eco Farm of Animal Science Faculty of Bogor Agricultural University (EcoFarm (20 hds; as well as from semi-intensive managements of small dairy farmers under the supervision of the North Lembang Cooperative Unit (KPSBU Lembang in two villages of Cilumber (98 hds and Pasar Kemis (92 hds. Blood samples were also taken via cocsigalis vein of AI (active and non active HF bulls at the two National AI Stations in Lembang (25 hds and Singosari (32 hds. Identification of genetic variation was by applying PCR-RFLP method. Three genotypes were identified, namely AA, AB, and BB, resulting two alleles of A and B. Results entirely showed that the frequencies of the BB HF female were very low, with the range of 0%-6% (vs AA genotype= 10%-54% and AB genotype= 46%-85%, despite of the relatively high frequency of the B allele over the A allele (23%-48% vs. 52%-77%. The low frequency of the observed BB females might be due to the limited AI active BB bulls used for services by the two national AI stations (0%-4%.

To compare the incidence of gastrointestinal side effects of two enteral feeding formulas with changes in the origin of protein (casein and isolated soy protein) in hospitalised elderly patients. A cross sectional survey was done among a sample of elderly patients carrying nasogastric tube admitted to the Reina Sofia General Hospital (Murcia) during a period of 6 months. A formula based on casein or soybean protein was randomly assigned. The variables studied were: age, sex, cause for indication of EN, duration of the EN and maximum amount of EN administered per day. Nutritional status at admission and discharge, mortality and gastrointestinal side effects (diarrhoea, constipation, vomits or regurgitation) were also collected. Statistical analyses were performed with the Student's T and chi 2 tests, with a significance of 95%. Sample conformed by 50 patients over 65 years (48% casein, 52% soybean) without statistically significant differences in age nor cause of indication of the EN. Either there were no differences in the nutritional status at the admission and discharge in both groups. Significant differences were observed in the incidence of diarrhoea (C: 45.83%, S: 7.69%, p = 0.009) and vomits (C: 41.66%, S: 15.38%, p = 0.05). A significant reduction in the incidence of gastrointestinal complications, a reduction in the incidence of ulcers by pressure and less mortality occurred on the group that took formula based on the soybean protein. The individualized nutritional evaluation must be performed routinely when the patient is admitted to the hospital for detection and treatment of early signs of malnutrition.

Casein kinase 1 epsilon (CK1ε) and its closest homologue CK1δ are key regulators of diverse cellular processes. We report two crystal structures of PF4800567, a potent and selective inhibitor of CK1ε, bound to the kinase domains of human CK1ε and CK1δ as well as one apo CK1ε crystal structure. These structures provide a molecular basis for the strong and specific inhibitor interactions with CK1ε and suggest clues for further development of CK1δ inhibitors.

Genetic polymorphisms of bovine milk proteins affect the protein profile of the milk and, hence, certain technological properties, such as casein (CN) number and cheese yield. However, reports show that such polymorphisms may also affect the health-related properties of milk. Therefore, to gain insight into their digestion pattern and bioactive potential, β-CN was purified from bovine milk originating from cows homozygous for the variants A(1), A(2), B, and I by a combination of cold storage, ultracentrifugation, and acid precipitation. The purity of the isolated β-CN was determined by HPLC, variants were verified by mass spectrometry, and molar extinction coefficients at λ=280nm were determined. β-Casein from each of the variants was subjected to in vitro digestion using pepsin and pancreatic enzymes. Antioxidant and angiotensin-converting enzyme (ACE) inhibitory capacities of the hydrolysates were assessed at 3 stages of digestion and related to that of the undigested samples. Neither molar extinction coefficients nor overall digestibility varied significantly between these 4 variants; however, clear differences in digestion pattern were indicated by gel electrophoresis. In particular, after 60min of pepsin followed by 5min of pancreatic enzyme digestion, one ≈4kDa peptide with the N-terminal sequence (106)H-K-E-M-P-F-P-K- was absent from β-CN variant B. This is likely a result of the (122)Ser to (122)Arg substitution in variant B introducing a novel trypsin cleavage site, leading to the changed digestion pattern. All investigated β-CN variants exhibited a significant increase in antioxidant capacity upon digestion, as measured by the Trolox-equivalent antioxidant capacity assay. After 60min of pepsin + 120min of pancreatic enzyme digestion, the accumulated increase in antioxidant capacity was ≈1.7-fold for the 4 β-CN variants. The ACE inhibitory capacity was also significantly increased by digestion, with the B variant reaching the highest inhibitory

The conformation of deamidated gliadins and b-casein in solution and adsorbed at the interface of oil-inwater emulsions was studied using synchrotron radiation circular dichroism (SRCD) and front-facefluorescence spectroscopy. Deamidation led to partial unfolding of gliadins in solution. The a....... In contrast, b-casein adopted a more ordered structure upon adsorption to these two oil/water interfaces, the a-helix content increased from 5.5% (in solution) to 20% and 22.5% respectively after adsorption to tricaprin/water and hexadecane/water interfaces. Both deamidated gliadins and b-casein have...... to oil/water interfaces. The hydrophobicity of the oil phase also has an impact on the conformation of each protein upon adsorption to the oil/water interfaces e systematic trends were observed between oil phase polarity from: i) tryptophan fluorescence emission maxima, and ii) the ahelix content...

The caseins comprise a milk protein fraction of high nutritional value and, as more recently discovered, of immunologic relevance. In particular, αS1-casein (CSN1S1) is of interest being a potential autoantigen. So far, the concentration of caseins in human milk was primarily determined by indirect methods. The aim of this study was to directly measure the CSN1S1 content in breast milk using mass spectrometry (MS). The quantification was based on tryptic CSN1S1 peptides with the best response in liquid chromatography (LC)-MS/MS analysis. Targeted experiments allowed both specific and sensitive detection at the low fmol level. For this pilot study, twenty breast milk samples of the first week post-partum were analyzed and contained between 3 and 540μg/ml CSN1S1. Limitations of CSN1S1 quantification are discussed.

When colloidal particles in a dispersion are made attractive, they aggregate into fractal clusters which grow to form a space-spanning network, or gel, even at low volume fractions. These gels are crucial to the rheological behavior of many personal care, food products and dispersion-based paints. The mechanical stability of these products relies on the stability of the colloidal gel network which acts as a scaffold to provide these products with desired mechanical properties and to prevent gravitational sedimentation of the dispersed components. Understanding the mechanical stability of such colloidal gels is thus of crucial importance to predict and control the properties of many soft solids. Once a colloidal gel forms, the heterogeneous structure bonded through weak physical interactions, is immediately subject to body forces, such as gravity, surface forces, such as adhesion to a container walls and shear forces; the interplay of these forces acting on the gel determines its stability. Even in the absence of external stresses, colloidal gels undergo internal rearrangements within the network that may cause the network structure to evolve gradually, in processes known as aging or coarsening or fail catastrophically, in a mechanical instability known as syneresis. Studying gel stability in the laboratory requires model colloidal system which may be tuned to eliminate these body or endogenous forces systematically. Using existing chemistry, I developed several systems to study delayed yielding by eliminating gravitational stresses through density matching and cyclic heating to induce attraction; and to study syneresis by eliminating adhesion to the container walls, altering the contact forces between colloids, and again, inducing gelation through heating. These results elucidate the varied yet concomitant mechanisms by which colloidal gels may locally or globally yield, but then reform due to the nature of the physical, or non-covalent, interactions which form

Responsive biomaterial hydrogels attract significant attention due to their biocompatibility and degradability. In order to make chitosan based gels, we first graft one layer of chitosan to silica, and then build a chitosan/poly(acrylic acid) multilayer using the layer-by-layer approach. After...... cross-linking the chitosan present in the polyelectrolyte multilayer, poly(acrylic acid) is partly removed by exposing the multilayer structure to a concentrated carbonate buffer solution at a high pH, leaving a surface-grafted cross-linked gel. Chemical cross-linking enhances the gel stability against......-linking density. The amount of poly(acrylic acid) trapped inside the surface grafted films was found to decrease with decreasing cross-linking density, as confirmed in situ using TIRR, and ex situ by Fourier transform infrared (FTIR) measurements on dried films. The responsiveness of the chitosan-based gels...

The flavonoid rutin is recognized as playing an important role in the protection of plants against lepidopterans. Bioassays with this compound are generally carried out using artificial diets. Proteins of high energy value, such as casein, are important ingredients of insect artificial diets as a source of essential amino acids. However, such proteins can generally increase the allelochemical activity. Our objective was to evaluate the effects of rutin on larvae of the velvetbean caterpillar Anticarsia gemmatalis Hübner by incorporating this allelochemical into diets with different concentrations of casein. Three casein concentrations (0, 7 g, or 14 g) combined with none, 0.65%, or 1.30% of rutin were added to the rearing diet and offered to the larvae from hatching to pupation. Rutin negatively affected larval development, the amount of food consumed, and pupal weight of A. gemmatalis. These negative effects were clearly seen in insects fed on diets with 7 g of casein to which any concentration of rutin was added. The effects of rutin when added to the diets without casein were stronger than in diets containing a suitable amount of casein (14 g). The greater negative effects of rutin in diets containing suboptimal concentrations of casein indicate that casein can increase the effects of rutin only when the diets are nutritionally unsuitable for insect development.

Full Text Available The effects of kappa-beta-casein genotypes and b-lactoglobulin genotypes on the renneting properties and composition of milk were estimated for 174 and 155 milk samples of 59 Finnish Ayrshire and 55 Finnish Friesian cows, respectively. As well as the random additive genetic and permanent environmental effects of a cow, the model included the fixed effects for parity, lactation stage, season, kappa-beta-casein genotypes and b-lactoglobulin genotypes. Favourable renneting properties were associated with kappa-beta-casein genotypes ABA 1 A 2 , ABA 1 A 1 and AAA 1 A 2 in the Finnish Ayrshire, and with ABA 2 B, AAA 1 A 3 , AAA 2 A 3 , ABA 1 A 2 and ABA 2 A 2 in the Finnish Friesian. The favourable effect of these genotypes on curd firming time and on firmness of the curd was partly due to their association with a high kappa-casein concentration in the milk. The effect of the kappa-casein E allele on renneting properties was unfavourable compared with that of the kappa-casein B allele, and possibly with that of the A allele. The beta-lactoglobulin genotypes had no effect on renneting properties but they had a clear effect on the protein composition of milk. The beta-lactoglobulin AA genotype was associated with a high whey protein % and beta-lactoglobulin concentration and the BB genotype with a high casein % and casein number.;

, as similar experiment with bovine serum albumin resulted in residual activity of horseradish peroxidase that was significantly lower than without any addition. The effect of β-casein on HRP disappears when pH is below the isoelectric point of β-casein. It was also proven by light scattering studies that β...

Nutritional interventions are important for reducing obesity and related conditions. Soy is a good source of protein and also contains isoflavones that may affect plasma lipids, body weight, and insulin action. Described here are data from a monkey breeding colony in which monkeys were initially fed a standard chow diet that is low fat with protein derived from soy. Monkeys were then randomized to a defined diet with a fat content similar to the typical American diet (TAD) containing either protein derived from soy (TAD soy) or casein-lactalbumin (TAD casein). The colony was followed for over two years to assess body weight, and carbohydrate and lipid measures in adult females (n=19) and their offspring (n=25). Serum isoflavone concentrations were higher with TAD soy than TAD casein, but not as high as when monkey chow was fed. Offspring consuming TAD soy had higher serum isoflavone concentrations than adults consuming TAD soy. Female monkeys consuming TAD soy had better glycemic control, as determined by fructosamine concentrations, but no differences in lipids or body weight compared with those consuming diets with TAD casein. Offspring born to dams consuming TAD soy had similar body weights at birth but over a two-year period weighed significantly less, had significantly lower triglyceride concentrations, and like adult females, had significantly lower fructosamine concentrations compared to TAD casein. Glucose tolerance tests in adult females were not significantly different with diet, but offspring eating TAD soy had increased glucose disappearance with overall lower glucose and insulin responses to the glucose challenge compared with TAD casein. Potential reasons for the additional benefits of TAD soy observed in offspring but not in adults may be related to higher serum isoflavone concentrations in offspring, presence of the diet differences throughout more of their lifespan (including gestation), or different tissue susceptibilities in younger animals.

The effects of heat stress on dairy production can be separated into 2 distinct causes: those effects that are mediated by the reduced voluntary feed intake associated with heat stress, and the direct physiological and metabolic effects of heat stress. To distinguish between these, and identify their effect on milk protein and casein concentration, mid-lactation Holstein-Friesian cows (n = 24) were housed in temperature-controlled chambers and either subjected to heat stress [HS; temperature-humidity index (THI) ~78] or kept in a THIheat-stressed cows (TN-R) for 7 d. A control group of cows was kept in a THIheat stress. Heat stress reduced the milk protein concentration, casein number, and casein concentration and increased the urea concentration in milk beyond the effects of restriction of intake. Under HS, the proportion in total casein of αS1-casein increased and the proportion of αS2-casein decreased. Because no effect of HS on milk fat or lactose concentration was found, these effects appeared to be the result of specific downregulation of mammary protein synthesis, and not a general reduction in mammary activity. No residual effects were found of HS or TN-R on milk production or composition after THIHeat-stressed cows had elevated blood concentrations of urea and Ca, compared with TN-R and TN-AL. Cows in TN-R had higher serum nonesterified fatty acid concentrations than cows in HS. It was proposed that HS and TN-R cows may mobilize different tissues as endogenous sources of energy.

Although Chou-Fasman calculations of the secondary structure of recombinant casein kinase 2 subunits alpha and beta suggest they have a similar overall conformation, circular dichroism (CD) studies show that substantial differences in the conformation of the two subunits exist. In addition......, no changes in the far-UV CD spectrum of the alpha subunit are observed in the presence of casein or the synthetic decapeptide substrate RRRDDDSDDD. Furthermore, the alpha-helical structure of the alpha subunit (but not the beta subunit) can be increased in the presence of stoichiometric amounts of heparin...

Hydrophobic silicon-based material having magnetic properties was fairly synthesized by a classical sol-gel approach. Pepsin enzyme was encapsulated in the sol-gel material and the enzyme activity was evaluated in consequence of the digestion of some common proteins such as α- and β-casein, cytochrome c, myoglobin, and bovine serum albumin (BSA) both in a single protein batch and in the protein mixture. The optimum digestion time of the studied proteins using pepsin-encapsulated magnetic sol-gel material was found to be 20min. To produce the magnetic sol-gel material for convenient and easy proteomics applications, Fe3O4 was doped inside sol-gel material during the gelation step. It was observed that the activity of encapsulated pepsin was not affected by the amount of Fe3O4. Poly(ethylene glycol) was also inserted in sol-gel bulk to obtain suitable roughness and increase the hydrophilicity of the material surface to let protein molecules reach to the sol-gel material easily. The digestion of the protein mixture and non-fat bovine milk was performed with the pepsin-encapsulated magnetic sol-gel material and the digested solutions were analyzed using SDS-PAGE, MALDI-TOF-MS and LC-MS/MS for the protein identification. Reusability of the pepsin-encapsulated sol-gel material was examined and it was determined that they could be used at least 20 times. Finally, IgG digestions with a fast incubation time period were carried out using pepsin-encapsulated sol-gel material for generation of (Fab)2 product to evaluate the kinetic performance of the material.

AIM: The aim of this conference paper was to systematically review the quality of evidence and summarize the findings of clinical trials published after 2002 using fluoride mouth rinses, fluoride gels or foams for the prevention of dental caries. METHODS: Relevant papers were selected after...... (6 on fluoride mouth rinse, 10 on fluoride gel and 3 on fluoride foam); 6 had a low risk of bias while 2 had a moderate risk. All fluoride measures appeared to be beneficial in preventing crown caries and reversing root caries, but the quality of evidence was graded as low for fluoride mouth rinse......, moderate for fluoride gel and very low for acidulated fluoride foam. No conclusions could be drawn on the cost-effectiveness. CONCLUSIONS: This review, covering the recent decade, has further substantiated the evidence for a caries-preventive effect of fluoride mouth rinse, fluoride gel and foam...

Empty liquids and equilibrium gels have so far been only theoretical possibilities, predicted for colloids with patchy interactions. But evidence of both has now been found in Laponite, a widely studied clay.

Full Text Available Profitable milk production respects the interests of producers, processing industries, consumer requirements and welfare of animals. Development of new methods of direct gene analysis responsible for milk proteins polymorphism provide new tools to raise the profitability of milk production and dairy products through implementation of breed genetic profile in breeding program. Because of necessity to determinate genetic profiles of cattle breeds in Croatia using new analytical methods, the ratio of dominant allelic polymorphic variants of beta-lactoglobulin (β-Lg and kappa-casein (κ-CN is defined. The share of beta-lactoglobulin B variant is dominant in all investigated cattle breeds (>52.9 %. Kappa- casein allelic variant A is dominant in selected cattle breeds (60.7-76.4 %, while the share of B variant is significantly more presented in autochthonous cattle breeds (48.2-84.1 %. Knowledge about genetic profile of breeds due to studied polymorphic variants of milk proteins is useful in further breeding development and economic reaffirmation of cattle breeds, especially autochthonous ones.

Proteins and polysaccharides are widely used in food formulation. While most of the proteins are surface active, only few polysaccharides can adsorb at the air-water interface; this is the case of propylene glycol alginates (PGA). It is known that casein glycomacropeptide (CMP), a bioactive polypeptide derived from κ-casein by the action of chymosin, presents a great foaming capacity but provides unstable foams. So, the objective of this work was to analyze the impact of mixing CMP and a commercial variety of PGA, Kelcoloid O (KO), on the interfacial and foaming properties at pH 7.0. It was determined the surface pressure isotherm, the dynamics of adsorption and the foaming properties for CMP, KO and the mixed system CMP-KO. CMP dominated the surface pressure of CMP-KO mixed system. The presence of KO synergistically improved the viscoelastic properties of surface film. The foaming capacity of CMP was altered by KO. KO foam presented a higher stability than CMP foam and it controlled the stability against drainage and the initial collapse in the mixed foam.

Many diet regimens were studied for patients with autism spectrum disorder (ASD) over the past few years. Ketogenic diet is gaining attention due to its proven effect on neurological conditions like epilepsy in children. Forty-five children aged 3-8 years diagnosed with ASD based on DSM-5 criteria were enrolled in this study. Patients were equally divided into 3 groups, first group received ketogenic diet as modified Atkins diet (MAD), second group received gluten free casein free (GFCF) diet and the third group received balanced nutrition and served as a control group. All patients were assessed in terms of neurological examination, anthropometric measures, as well as Childhood Autism Rating Scale (CARS), Autism Treatment Evaluation Test (ATEC) scales before and 6 months after starting diet. Both diet groups showed significant improvement in ATEC and CARS scores in comparison to control group, yet ketogenic scored better results in cognition and sociability compared to GFCF diet group. Depending on the parameters measured in our study, modified Atkins diet and gluten free casein free diet regimens may safely improve autistic manifestations and could be recommended for children with ASD. At this stage, this study is a single center study with a small number of patients and a great deal of additional wide-scale prospective studies are however needed to confirm these results. UMIN-CTR Study Design: trial Number UMIN000021433.

Casein kinase 1 delta (CK1δ) and casein kinase 1 epsilon (CK1ε) inhibitors are potential therapeutic agents for a range of psychiatric disorders. The feasibility of developing a CNS kinase inhibitor has been limited by an inability to identify safe brain-penetrant compounds with high kinome selectivity. Guided by structure-based drug design, potent and selective CK1δ/ε inhibitors have now been identified that address this gap, through the design and synthesis of novel 4-[4-(4-fluorophenyl)-1-(piperidin-4-yl)-1H-imidazol-5-yl]pyrimidin-2-amine derivatives. PF-5006739 (6) possesses a desirable profile, with low nanomolar in vitro potency for CK1δ/ε (IC50 = 3.9 and 17.0 nM, respectively) and high kinome selectivity. In vivo, 6 demonstrated robust centrally mediated circadian rhythm phase-delaying effects in both nocturnal and diurnal animal models. Further, 6 dose-dependently attenuated opioid drug-seeking behavior in a rodent operant reinstatement model in animals trained to self-administer fentanyl. Collectively, our data supports further development of 6 as a promising candidate to test the hypothesis of CK1δ/ε inhibition in treating multiple indications in the clinic.

The COMMA-D mammary cell line exhibits mammary-specific functional differentiation under appropriate conditions in cell culture. The cytologically heterogeneous COMMA-D parental line and the clonal lines DB-1, TA-5, and FA-1 derived from the COMMA-D parent were examined for similar properties of functional differentiation. In monolayer cell culture, the cell lines DB-1, TA-5, FA-1, and MA-4 were examined for expression of mammary-specific and epithelial-specific proteins by an indirect immunofluorescence assay. The clonal cell lines were relatively homogeneous in their respective staining properties and seemed to represent three subpopulations found in the heterogeneous parental COMMA-D lines. None of the four clonal lines appeared to represent myoepithelial cells. The cell lines were examined for expression of {beta}-casein mRNA in the presence or absence of prolactin. The inducibility of {beta}-casein in the COMMA-D cell line was further enhanced by a reconstituted basement membrane preparation enriched in laminin, collagen IV, and proteoglycans. These results support the hypothesis that the functional response of inducible mammary cell populations is a result of interaction among hormones, multiple extracellular matrix components, and specific cell types.

Macroscopic properties and ensemble average diffusion of concentrated (dispersed phase 50-60 wt%) Na-caseinate-stabilised emulsions for three different oils (soybean oil, palm olein and tetradecane) were explored. On a volume fraction basis, pulsed gradient stimulated echo (PGSTE)-NMR data show that droplet dynamics for all three systems are similar within a region of the emulsion morphology diagram. The exact limits of the emulsion space depend however on which oil is considered. The reduced solubility of tetradecane in water, and Na-caseinate in tetradecane, result in the stabilisation of flocs during formulation. Floc formation is not observed when soybean oil or palm olein is used under identical emulsion formulation conditions. Linear rheology experiments provide indirect evidence that the local structure and the properties of the thin film interfacial domain of tetradecane emulsions vary from those of soybean oil and palm olein emulsions. Collectively these data indicate that protein/oil interactions within a system dominate over specific oil droplet structure and size distribution, which are similar in the three systems.

We have demonstrated a solution-based synthesis of novel waxberry-like hierarchical ZnO microparticles in the presence casein micelles under mild conditions. The microstructures of the sub-micrometer-sized hierarchical microparticles were characterized, and the synthesis conditions were optimized. The formation mechanism of the hierarchical microparticle was analyzed through control experiments. The hierarchical ZnO microparticles are found to be super-assemblies of 30-70 nm ZnO nanoparticles, which are thought to be based on casein micelle induction followed by Ostwald ripening. In the same manner, copper-based hierarchical microparticles with a similar morphology have also been successfully synthesized. By controlling the synthetic time or temperature, solid or hollow microparticles can be fabricated. The narrowly distributed ZnO microparticles have a high specific surface area, exhibiting great potential application in fields such as photocatalytic and energy conversion. Our findings may meanwhile open a new bottom-up strategy in order to construct structurally sophisticated nanomaterials.

The impact of a cationic surfactant, dodecylammonium chloride (DDACl), on the self-assembly of sodium caseinate (SC) has been investigated by light scattering, zeta potential, and rheological measurements as well as by microscopy (transmission electron and confocal laser scanning microscopy). In SC dilute solutions concentration-dependent self-assembly proceeds through the formation of spherical associates and their aggregation into elongated structures composed of connected spheres. DDACl interacts with SC via its hydrophilic and hydrophobic groups, inducing changes in SC self-assembled structures. These changes strongly depend on the surfactant aggregation states (monomeric or micellar) as well as concentration ratio of both components, leading to the formation of soluble and insoluble complexes of nano- to microdimensions. DDACl monomers interact with SC self-assembled entities in a different way compared to their micelles. Surfactant monomers form soluble complexes (similar to surfactant mixed micelles) at lower SC concentration but insoluble gelatinous complexes at higher SC concentration. At surfactant micellar concentration soluble complexes with casein chains wrapped around surfactant micelles are formed. This study suggests that the use of proper cationic surfactant concentration will allow modification and control of structural changes of SC self-assembled entities.

This study aims to investigate the effect of topical applications of 10% casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) on white spot lesions (WSL) detected after treatment with fixed orthodontic appliances. Sixty healthy adolescents with ≥1 clinically visible WSL at debonding were recruited and randomly allocated to a randomised controlled trial with two parallel groups. The intervention group was instructed to topically apply a CPP-ACP -containing agent (Tooth Mousse, GC Europe) once daily and the subjects of the control group brushed their teeth with standard fluoride toothpaste. The intervention period was 4 weeks and the endpoints were quantitative light-induced fluorescence (QLF) on buccal surfaces of the upper incisors, cuspids and first premolars and visual scoring from digital photos. The attrition rate was 15%, mostly due to technical errors, and 327 lesions were included in the final evaluation. A statistically significant (p white spot lesions after debonding of orthodontic appliances with a casein phosphopeptide-stabilised amorphous calcium phosphate agent resulted in significantly reduced fluorescence and a reduced area of the lesions after 4 weeks as assessed by QLF. The improvement was however not superior to the "natural" regression following daily use of fluoride toothpaste.

Full Text Available Background: Even though the etiology of autistic spectrum disorder (ASD has not been clearly known, various types of therapies have been offered. One of the most popular therapies is Gluten Free and Casein Free Diet – GFCF diet.Methods: A structured internet literature search was conducted using Pubmed and Cochrane library. Titles and abstracts were screened using predetermined selection criteria. Critical appraisal was conducted based on standard criteria for relevance, validity, and levels of evidence.Results: Based on relevance, only four out of twelve selected articles can be used; one is a systematic review and three randomized controlled trials (RCTs. Several methodological problems which could lead to over-estimation of the effect were found in the studies including poor randomization and selective drop-out. Risk of under-estimation of report due to small number of participants and in-adequate duration of intervention was also found. Small to moderate improvement were found in some outcome measured including overall autistic traits, social isolation, and the ability in communication and interaction. However, insignificant differences also found in some area such as individual’s behavior, cognitive and motor function.Conclusion: Available studies fail to provide sufficiently credible and strong evidence to recommend the practice of GFCF diet in treating autism. (Med J Indones 2011; 20:114-8Keywords: autism, gluten free and casein free diet

Precise contact between epithelial cells and their underlying basement membrane is critical to the maintenance of tissue architecture and function. To understand the role that the laminin receptor dystroglycan (DG) plays in these processes, we assayed cell responses to laminin-111 following conditional ablation of DG expression in cultured mammary epithelial cells (MECs). Strikingly, DG loss disrupted laminin-111-induced polarity and {beta}-casein production, and abolished laminin assembly at the step of laminin binding to the cell surface. DG re-expression restored these deficiencies. Investigations of mechanism revealed that DG cytoplasmic sequences were not necessary for laminin assembly and signaling, and only when the entire mucin domain of extracellular DG was deleted did laminin assembly not occur. These results demonstrate that DG is essential as a laminin-111 co-receptor in MECs that functions by mediating laminin anchoring to the cell surface, a process that allows laminin polymerization, tissue polarity, and {beta}-casein induction. The observed loss of laminin-111 assembly and signaling in DG-/-MECs provides insights into the signaling changes occurring in breast carcinomas and other cancers, where DG's laminin-binding function is frequently defective.

Very high casein content and good coagulation properties previously observed in some Ethiopian goat breeds led to investigating the αs1-casein (CSN1S1) gene in these breeds. Selected regions of the CSN1S1 gene were sequenced in 115 goats from 5 breeds (2 indigenous: Arsi-Bale and Somali, 1 exotic: Boer, and 2 crossbreeds: Boer × Arsi-Bale and Boer × Somali). The DNA analysis resulted in 35 new mutations: 3 in exons, 3 in the 5' untranslated region (UTR), and 29 in the introns. The mutations in exons that resulted in an amino acid shift were then picked to evaluate their influence on individual casein content (αs1-, αs2-, β-, and κ-CN), micellar size, and coagulation properties in the milk from the 5 goat breeds. A mutation at nucleotide 10657 (exon 10) involved a transversion: CAG→CCG, resulting in an amino acid exchange Gln77→Pro77. This mutation was associated with the indigenous breeds only. Two new mutations, at nucleotide 6072 (exon 4) and 12165 (exon 12), revealed synonymous transitions: GTC→GTT in Val15 and AGA→AGG in Arg100 of the mature protein. Transitions G→A and C→T at nucleotides 1374 and 1866, respectively, occurred in the 5' UTR, whereas the third mutation involved a transversion T→G at nucleotide location 1592. The goats were grouped into homozygote new (CC), homozygote reference (AA), and heterozygote (CA) based on the nucleotide that involved the transversion. The content of αs1-CN (15.32g/kg) in milk samples of goats homozygous (CC) for this newly identified mutation, Gln77→Pro77 was significantly higher than in milks of heterozygous (CA; 9.05g/kg) and reference (AA; 7.61g/kg) genotype animals. The αs2-, β-, and κ-CN contents showed a similar pattern. Milk from goats with a homozygous new mutation had significantly lower micellar size. Milk from both homozygote and heterozygote new-mutation goats had significantly shorter coagulation rate and stronger gel than the reference genotype. Except the transversion, the

The effect of silica and polymer coated silica particles as reinforcing agents on the structural and mechanical properties of polystyrene-poly(ethylene/butylene)-polystyrene (PS-PEB-PS) triblock gel has been investigated. Different types of chemically modified silica have been compared in order...... a viscoclastic rubber to a plastic fluid and from a plastic fluid to a viscoelastic liquid are shifted to more elevated temperatures when silica is added to the triblock copolymer gel. (C) 2004 Elsevier Ltd. All rights reserved....

Casein phosphorylation is a posttranslational modification catalyzed by kinase enzymes that attach phosphate groups to specific AA in the protein sequence. This modification is one of the key factors responsible for the stabilization of calcium phosphate nanoclusters in casein micelles and for the internal structure of the casein micelles. α(S1)-Casein (α(s1)-CN) is of special interest because it constitutes up to 40% of the total casein fraction in milk, and it has 2 common phosphorylation states, with 8 (α(S1)-CN-8P) and 9 (α(S1)-CN-9P) phosphorylated serine residues. Factors affecting this variation in the degree of phosphorylation are not currently known. The objective of this research was to determine the genetic background of α(S1)-CN-8P and α(S1)-CN-9P. The genetic and phenotypic correlation between α(S1)-CN-8P and α(S1)-CN-9P was low (0.18 and 0.19, respectively). This low genetic correlation suggests a different genetic background. These differences were further investigated by means of a genome-wide association study, which showed that both α(S1)-CN-8P and α(S1)-CN-9P were affected by a region on Bos taurus autosome (BTA) 6, but only α(S1)-CN-8P was affected by a region on BTA11 that contains the gene that encodes for β-lactoglobulin (β-LG), and only α(S1)-CN-9P was affected by a region on BTA14 that contains the diacylglycerol acyltransferase 1 (DGAT1) gene. Estimated effects of β-LG protein genotypes showed that only α(S1)-CN-8P was associated with the β-LG A/B polymorphism (g.1772G>A and g.3054C>T); the AA genotype of β-LG was associated with a lower concentration of α(S1)-CN-8P (-0.32% wt/wt) than the BB genotype (+0.41% wt/wt). Estimated effects of DGAT1 K232A genotypes showed that only α(S1)-CN-9P was associated with the DGAT1 gene polymorphism; DGAT1 AA genotype was associated with a higher α(S1)-CN-9P concentration (+0.53% wt/wt) than the DGAT1 KK genotype (-0.44% wt/wt). The results give insight in phosphorylation of α(S1

Responsive biomaterial hydrogels attract significant attention due to their biocompatibility and degradability. In order to make chitosan based gels, we first graft one layer of chitosan to silica, and then build a chitosan/poly(acrylic acid) multilayer using the layer-by-layer approach. After cross-linking the chitosan present in the polyelectrolyte multilayer, poly(acrylic acid) is partly removed by exposing the multilayer structure to a concentrated carbonate buffer solution at a high pH, leaving a surface-grafted cross-linked gel. Chemical cross-linking enhances the gel stability against detachment and decomposition. The chemical reaction between gluteraldehyde, the cross-linking agent, and chitosan was followed in situ using total internal reflection Raman (TIRR) spectroscopy, which provided a molecular insight into the complex reaction mechanism, as well as the means to quantify the cross-linking density. The amount of poly(acrylic acid) trapped inside the surface grafted films was found to decrease with decreasing cross-linking density, as confirmed in situ using TIRR, and ex situ by Fourier transform infrared (FTIR) measurements on dried films. The responsiveness of the chitosan-based gels with respect to pH changes was probed by quartz crystal microbalance with dissipation (QCM-D) and TIRR. Highly cross-linked gels show a small and fully reversible behavior when the solution pH is switched between pH 2.7 and 5.7. In contrast, low cross-linked gels are more responsive to pH changes, but the response is fully reversible only after the first exposure to the acidic solution, once an internal restructuring of the gel has taken place. Two distinct pKa's for both chitosan and poly(acrylic acid), were determined for the cross-linked structure using TIRR. They are associated with populations of chargeable groups displaying either a bulk like dissociation behavior or forming ionic complexes inside the hydrogel film.

subunits are highly conserved during evolution. The relationship between CK-2 alpha from humans and plants is still 73%. Similar relationships are reported for the beta-subunit. Chromosomal assignment of CK-2 alpha shows two gene loci, one of which is a pseudogene. They are located on different chromosomes......, no genetic changes are necessarily involved; the observed changes may be entirely due to a signal transduction pathway where CK-2 could be phosphorylated by another kinase(s). CK-2 cDNAs from various organisms have been isolated and characterized. From the deduced amino acid sequence it turns out that CK-2......-subunit affecting: (i) stability, (ii) enzyme specificity and (iii) enzyme activity. The question where CK-2 and its subunits are located throughout the cell cycle has also been addressed, mainly because of the large discrepancies that still exist between results obtained by different investigators. Tissue...

Full Text Available Aim: This study aims to determine and compare the extent of inhibition of demineralization and promotion of remineralization of permanent molar enamel with and without application of three remineralizing agents. Materials and Methods: Forty extracted permanent molars were randomly divided into two groups 1 and 2, longitudinally sectioned into four and divided into subgroups A, B, C, and D. The sections were coated with nail varnish leaving a window of 3 mm × 3 mm. All sections of Group 1 were treated with their respective subgroup-specific agent: Casein phosphopeptide-amorphous calcium phosphate (CPP-ACP paste for subgroup A, CPP-amorphous calcium phosphate fluoride (ACPF paste for subgroup B, CPP-ACPF varnish for subgroup C and subgroup D served as a control. The sections were then subjected to demineralization for 12 days following which lesional depth was measured under the stereomicroscope. All the sections of Group 2 were subjected to demineralization for 12 days, examined for lesional depth, then treated with their respective subgroup specific agents and immersed in artificial saliva for 7 days. The sections were then examined again under the stereomicroscope to measure the lesional depth. Results: CPP-ACPF varnish caused significant inhibition of demineralization. All three agents showed significant remineralization of previously demineralized lesions. However, CPP-ACPF varnish showed the greatest remineralization, followed by CPP-ACPF paste and then CPP-ACP paste. Conclusion: This study shows that CPP-ACPF varnish is effective in preventing demineralization as well as promoting remineralization of enamel. Thus, it can be used as an effective preventive measure for pediatric patients where compliance with the use of tooth mousse may be questionable.

We compared anthropometric values, nutrient intake, the Healthy Eating Index and food variety in children with autism spectrum disorder (ASD), 20 on a gluten-free casein-free (GFCF) diet and 85 on a regular diet in Valencia (Spain) using 3-days food diaries. Those on the GFCF diet had a lower weight, body mass index, and total energy, pantothenic…

Milk proteins were hydrolyzed by papain and their effect on the rheological, textural and sensory properties of bread were investigated. Water absorption capacity, emulsification capacity, foam volume, foam stability and solubility of Whey and casein protein concentrates and their hydrolysates were determined. The farinograph parameters of wheat flour and blends of wheat flour with casein and whey protein and their hydrolysates were determined to evaluate changes in water absorption capacity, dough development time, dough stability time and mixing tolerance index. The incorporation of WPC, casein and their hydrolysates up to the level of 5 % showed dough properties comparable to control. It was also found that 5 % level incorporation of milk proteins and their hydrolysates have no drastic effect on physical and sensory attributes of bread. The pasting properties showed significant decrease (p ≤ 0.05) when compared with wheat flour at all levels of addition of whey and casein protein concentrates and hydrolysates. Scanning electron microscopy of bread samples shows disruption in the well-defined protein - starch complex of wheat flour bread and the structure of gluten was weak as the concentration of whey protein increases in the wheat flour bread.

In practice it is challenging to prepare a concentrated medical product with high heat stability and low viscosity. Calcium chelators are often added to dairy products to improve heat stability, but this may increase viscosity through interactions with the casein proteins. The aim of

Recombinant human alpha subunit from casein kinase-2 (CK-2) was subjected, either alone or in combination with recombinant human beta subunit, to high temperature, tryptic digestion and urea treatment. In all three cases, it was shown that the presence of the beta subunit could drastically reduce...

In the present studies, several hypotheses were tested to explain previously reported differential effects of soy and casein on colon cancer biomarkers like cell proliferation, fecal fat, fecal bile acid, alkaline phosphatase, and magnesium excretion in rats. In Study 1, the effect of methionine, a

The present study examined whether different proteins have different effects on whole-body protein turnover in adult rats. The rats were either starved, given a protein-free but energy-sufficient diet (1 MJ/kg body weight (BW) per d) or a diet containing intact casein, hydrolysed casein, or hydro......The present study examined whether different proteins have different effects on whole-body protein turnover in adult rats. The rats were either starved, given a protein-free but energy-sufficient diet (1 MJ/kg body weight (BW) per d) or a diet containing intact casein, hydrolysed casein......, or hydrolysed soya-bean protein at a level of 9.1 g/kg BW per d. The diets, which were isoenergetic with the same carbohydrate: fat ratio, were given as a continuous intragastric infusion for at least 4 d. During the last 19 h 15N-glycine (a primed continuous infusion) was given intragastrically and 15N...... proteins was reflected in plasma amino acid composition and this is suggested to be responsible for the different effect on protein turnover. Preliminary results from this study have previously been published in abstract form (Nielsen et al. 1991)....

Casein kinase 1 (CK1) members play key roles in numerous biological processes. They are considered "rogue" kinases, because their enzymatic activity appears unregulated. Contrary to this notion, we have identified the DEAD-box RNA helicase DDX3 as a regulator of the Wnt-beta-catenin network, where i

This paper systematically reviews research on the effects of gluten-free and/or casein-free (GFCF) diets in the treatment of ASD. Database, hand, and ancestry searches identified 15 articles for review. Each study was analyzed and summarized in terms of (a) participants, (b) specifics of the intervention, (c) dependent variables, (d) results, and…

Whey protein has been demonstrated to improve fasting lipid and insulin response in overweight and obese individuals. To establish new hypotheses for this effect and to investigate the impact of stomach emptying, we compared plasma profiles after intake of whey isolate (WI), casein, gluten (GLU...

Shortly after the publication of our literature review on gluten-free and casein-free (GFCF) diets in the treatment of autism spectrum disorders (ASD; Mulloy et al., 2010), Whiteley et al. (2010) published the results of experimental evaluation of a GFCF diet. To update our previous literature review, we herein provide an analysis of the Whiteley…

This paper systematically reviews research on the effects of gluten-free and/or casein-free (GFCF) diets in the treatment of ASD. Database, hand, and ancestry searches identified 15 articles for review. Each study was analyzed and summarized in terms of (a) participants, (b) specifics of the intervention, (c) dependent variables, (d) results, and…

Shortly after the publication of our literature review on gluten-free and casein-free (GFCF) diets in the treatment of autism spectrum disorders (ASD; Mulloy et al., 2010), Whiteley et al. (2010) published the results of experimental evaluation of a GFCF diet. To update our previous literature review, we herein provide an analysis of the Whiteley…

Casein from bovine milk is evaluated in this work as binding agent for electrochemical double layer capacitors (EDLCs) electrodes. It is demonstrated that casein provides excellent adhesion strength to the current collector (1187 kPa compared to 51 kPa achieved with PVdF), thus leading to mechanically stable electrodes. At the same time, it offers high thermal stability (above 200 °C) and electrochemical stability in organic electrolytes. Apparently though, the casein-based electrodes offer lower electronic conductivity than those based on other state-of-the-art binders, which can limit the rate performance of the resulting EDLC. In the attempt of improving the electrochemical performance, it is found that the application of a pressing step can solve this issue, leading to excellent rate capability (up to 84% capacitance retention at 50 mA cm-2) and cycling stability (96.8% after 10,000 cycles at 10 mA cm-2) in both PC- and ACN-based electrolytes. Although the adhesive power casein is known since ancient times, this report presents the first proof of concept of its employment in electrochemical power sources.

In practice it is challenging to prepare a concentrated medical product with high heat stability and low viscosity. Calcium chelators are often added to dairy products to improve heat stability, but this may increase viscosity through interactions with the casein proteins. The aim of thi

Casein from ovine and bovine milk were hydrolysed with two extracellular protease preparations from Lactobacillus brevis and Lactococcus lactis. The hydrolysates were analyzed by HPLC-MS/MS for peptide identification. A strain-dependent peptide profile could be observed, regardless of the casein origin, and the specificity of these two proteases could be computationally ascribed. The cleavage pattern yielding phenylalanine, leucine, or tyrosine at C-terminal, appeared both at L. lactis and Lb. brevis hydrolysates. However, the cleavage C-terminal to lysine was favored with Lb. brevis protease. The hydrolysates showed ACE-inhibitory activity with IC50 in the 16-70 μg/ml range. Ovine casein hydrolysates yielded greater ACE-inhibitory activity. Previously described antihypertensive and opioid peptides were found in these ovine and bovine casein hydrolysates and prediction of the antihypertensive activity of the sequences based on quantitative structure and activity relationship (QSAR) was performed. This approach might represent a useful classification tool regarding health-related properties prior to further purification.

AFM measurements in the force volume mode were performed over the total penetration depth for different positions on casein aggregates adsorbed to a graphite surface in a liquid cell. The stiffness of the force curves was correlated to indentation depths, layer depth and lateral position within the

This study tested the efficacy of a gluten-free and casein-free (GFCF) diet in treating autism using a randomized, double blind repeated measures crossover design. The sample included 15 children aged 2-16 years with autism spectrum disorder. Data on autistic symptoms and urinary peptide levels were collected in the subjects' homes over the 12…

We investigated the effect of altering temperature immediately after gels were formed at 37 degrees C. We defined instrumentally measurable gelation (IMG) as the point at which gels had a storage modulus (G') > or = 5Pa. Gels were made at constant incubation temperature (IT) of 37 degrees C up to IMG, and then cooled to 30 or 33.5, or heated to 40.5 or 44 degrees C, at a rate of 1 degrees C/min and maintained at those temperatures until pH 4.6. Control gel was made at 37 degrees C (i.e., no temperature change during gelation/gel development). Gel formation was monitored using small strain dynamic oscillatory rheology, and the resulting structure and physical properties at pH 4.6 were studied by fluorescence microscopy, large deformation rheology, whey separation (WS), and permeability (B). A single strain of Streptococcus thermophilus was used to avoid variations in the ratios of strains that could have resulted from changes in temperature during fermentation. Total time required to reach pH 4.6 was similar for samples made at constant IT of 37 degrees C or by cooling after IMG from 37 to either 30 or 33.5 degrees C, but gels heated to 40 or 44 degrees C needed less time to reach pH 4.6. Cooling gels after IMG resulted in an increase in G' values at pH 4.6, a decrease in LT(max), WS, and B, and an increase in the area of protein aggregates of micrographs compared with the control gel made at constant IT of 37 degrees C. Heating gels after IMG resulted in a decrease in G' values at pH 4.6 and an increase in LT(max) values and WS. The physical properties of acid milk gels were dominated by the temperature profile during the gel-strengthening phase that occurs after IMG. This study indicates that the final properties of yogurt greatly depend on the environmental conditions (e.g., temperature, time/rate of pH change) experienced by the casein particles/clusters during the critical early gel development phase when bonding between and within particles is still labile

A thixotropic gel suitable for use in subsurface bioremediation is provided along with a process of using the gel. The thixotropic gel provides a non-migrating injectable substrate that can provide below ground barrier properties. In addition, the gel components provide for a favorable environment in which certain contaminants are preferentially sequestered in the gel and subsequently remediated by either indigenous or introduced microorganisms.

A thixotropic gel suitable for use in subsurface bioremediation is provided along with a process of using the gel. The thixotropic gel provides a non-migrating injectable substrate that can provide below ground barrier properties. In addition, the gel components provide for a favorable environment in which certain contaminants are preferentially sequestered in the gel and subsequently remediated by either indigenous or introduced microorganisms.

A thixotropic gel suitable for use in subsurface bioremediation is provided along with a process of using the gel. The thixotropic gel provides a non-migrating injectable substrate that can provide below ground barrier properties. In addition, the gel components provide for a favorable environment in which certain contaminants are preferentially sequestered in the gel and subsequently remediated by either indigenous or introduced microorganisms.

We present a statistical mechanical approach to predicting the properties of mixed copolymer layers using the Scheutjens-Fleer self-consistent-field theory. Our model copolymers are based on the primary structures of the major bovine casein monomers, alpha(s1)-casein and beta-casein. Numerical calculations have been carried out to determine the polymer segment density profiles at an isolated hydrophobic surface and the interaction forces as a pair of polymer-coated surfaces is brought to close interlayer separation. For a copolymer model containing hydrophilic and hydrophobic segments, we show how the steric stabilizing capacity of a casein-like macromolecule at very low surface coverage is enhanced in the presence of a thin dense layer of shorter tethered amphiphilic chains. Using a more refined protein model, which also distinguishes between the charged and uncharged hydrophilic segments along the chain, we clearly demonstrate that the enhanced steric repulsion from beta-casein exceeds that from alpha(s1)-casein. These calculations explain how the replacement of just a few percent of beta-lactoglobulin by casein can inhibit the heat-induced thickening and flocculation behavior observed experimentally with some whey protein-stabilized oil-in-water emulsions.

The hypothesis tested was that the inhibitory effect of dietary soy protein versus casein on fat digestion in veal calves would be smaller when diets were fed with high instead of low calcium content. Male calves, 1 wk of age, were fed 1 of 4 experimental milk replacers in a 2 x 2 factorial design. There were 19 animals per dietary group. The milk replacers contained either casein or soy protein isolate as variable protein source and were either low or high in calcium. Body weight gain was not significantly affected by the experimental diets. Soy protein isolate versus casein significantly reduced apparent fat digestibility. High versus low calcium intake also depressed fat digestion. The protein effect was smaller (2.9% units) for the high than the low calcium diets (3.6% units), but the interaction did not reach statistical significance. Soy protein isolate versus casein raised fecal bile acid excretion and so did high versus low calcium intake. The difference in bile acid excretion between the soy and casein containing diets was significantly greater for the high than low calcium diets. The absorption of phosphorus, calcium, and magnesium was higher for the casein diets than for the soy-containing diets. This study shows for the first time that soy protein isolate versus casein depressed fat digestion and raised fecal bile acid excretion in veal calves.

Full Text Available Thin casein films for food packaging applications reportedly possess good strength and low oxygen permeability, but low elasticity and high sensitivity to moisture. Modifying the films to target specific behaviors depending on environmental conditions can enable a variety of commercial applications for casein-based films. The mechanical properties of solvent-cast (15% solids calcium-caseinate/glycerol films (CaCas:Gly ratio of 3:1 were characterized as a function of processing and environmental conditions, including film thickness, solution formulation and ambient humidity (from 22% to 70% relative humidity (RH at ~20 °C. At constant RH, the elongation at break (EAB had a strong positive dependence on the film thickness. When RH increased, the tensile strength (TS and modulus (E decreased approximately linearly, while EAB increased. From 0.05% to 1% (w/w of citric pectin (CP was then incorporated into CaCas/Gly films following seven different formulations (mixing sequences, to alter the protein network and to evaluate the effects of CP on the tensile properties of CaCas/Gly/CP films. At constant film thickness and ~60% RH, the addition of 0.1% or 1.0% CP to the films considerably increased or decreased EAB, TS and E in different directions and to different extents, depending on the formulation, while optical micrographs also showed vastly differing network configurations, suggesting complex formulation- and stoichiometry-dependent casein-pectin interactions within the dried films. Depending on the desired film properties and utilization conditions, pectin may be a useful addition to casein film formulations for food packaging applications.

Casein-derived peptides are shown to possess radical scavenging and metal chelating properties. The objective of this study was to evaluate novel anti-inflammatory properties of casein hydrolysates (CH) produced by an eco-friendly process that combines high hydrostatic pressure with enzymatic hydrolysis (HHP-EH). Casein was hydrolysed by different proteases, including flavourzyme (Fla), savinase (Sav), thermolysin (Ther), trypsin (Try), and elastase (Ela) at 0.1, 50, 100, and 200 MPa pressure levels under various enzyme-to-substrate ratios and incubation times. Casein hydrolysates were evaluated for the degree of hydrolysis (DH), molecular weight distribution patterns, and anti-inflammatory properties in chemical and cellular models. Hydrolysates produced using HHP-EH exhibited higher DH values and proportions of smaller peptides compared to atmospheric pressure-enzymatic hydrolysis (AP-EH). Among five enzymes, Fla-digested HHP-EH-CH (HHP-Fla-CH) showed significantly higher antioxidant properties than AP-Fla-CH. The anti-inflammatory properties of HHP-Fla-CH were also observed by significantly reduced nitric oxide and by the suppression of the synthesis of pro-inflammatory cytokines in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophage cells. Liquid chromatography with tandem mass spectrometry (LC-MS/MS) revealed that 59% of the amino acids of the peptides in HHP-Fla-CH were composed of proline, valine, and leucine, indicating the potential anti-inflammatory properties. In conclusion, the HHP-EH method provides a promising technology to produce bioactive peptides from casein in an eco-friendly process.

The objective of this study was to estimate the effect of composite β-κ-CN genotypes on milk coagulation and composition traits, and on the additive genetic variation of these traits in Estonian Holstein dairy cattle. A total of 23,970 milk samples, repeated measurements from the first to third lactation from 2,859 Estonian Holstein cows from 78 herds across the country, were analyzed for milk yield, milk fat and protein percentages, somatic cell count, and milk coagulation properties (milk coagulation time and curd firmness). Each cow had at least 3 measurements per lactation. Two single-trait random regression animal models were fitted for the traits studied. The first model considered fixed effects of year-season of sampling and year-season of calving, calving age (nested within lactation), sample age (only for milk coagulation traits) and days in milk, and random herd, additive genetic, and permanent environmental effects. The animal and permanent environmental effects were modeled over the lactation period by using Legendre polynomials. The second model had the additional fixed β-κ-casein effect in the form of a third-order Legendre polynomial. The 2 most frequent β-κ-casein composite genotypes were A2A2AA and A1A2AA, both with prevalence greater than 20%. Percentages of the remaining 31 genotypes were less than 8%, including 20 genotypes with percentages less than 1%. The β-κ-casein genotype-specific lactation curves were significantly different for milk coagulation traits and milk protein percentage. The B variant of κ-casein showed a favorable effect on both milk coagulation traits, whereas the IB haplotype had an increasing effect on curd firmness and protein percentage. Inclusion of the β-κ-casein genotype effects in the model resulted in decreases in the mean additive genetic variations for milk coagulation time and curd firmness of 12.9 and 51.1%, respectively.

Full Text Available Normal 0 21 false false false PT-BR X-NONE X-NONE Solid lipid microparticles produced with a mixture of cupuacu butter and stearic acid were used to microencapsulate a commercial casein hydrolysate (Hyprol 8052. The composition of the lipid matrix used for the production of the lipid microparticles was chosen according to data on the wide angle X-ray diffraction (WAXD and differential scanning calorimetry (DSC of bulk lipid mixtures, which indicated that the presence of 10 % cupuacu butter was sufficient to significantly change the crystalline arrangement of pure stearic acid. Preliminary tests indicated that a minimum proportion of 4 % of surfactant (polysorbate 80 was necessary to produce empty spherical lipid particles with average diameters below 10 mm. The lipid microparticles were produced using 20 % cupuacu butter and 80 % stearic acid and then stabilized with 4 % of polysorbate 80, exhibiting an encapsulation efficiency of approximately 74 % of the casein hydrolysate. The melting temperature of the casein hydrolysate-loaded lipid microparticles was detected at 65.2 °C, demonstrating that the particles were solid at room temperature as expected and indicating that the incorporation of peptides had not affected their thermal behavior. After 25 days of storage, however, there was a release of approximately 30 % of the initial amount of encapsulated casein hydrolysate. This release was not thought to have been caused by the liberation of encapsulated casein hydrolysate. Instead, it was attributed to the possible desorption of the adsorbed peptides present on the surface of the lipid microparticles.

Six duodenally and ileally cannulated steers were used in 3 sequential studies to measure 1) basal nutrient flows from a soybean hull-based diet, 2) small intestinal digestibility of raw cornstarch continuously infused into the duodenum, and 3) responses of small intestinal starch digestion to duodenal infusion of 200 or 400 g/d casein. Our objective was to evaluate responses in small intestinal starch digestion in cattle over time and to measure responses in small intestinal starch digestion to increasing amounts of MP. On average, cattle consumed 3.7 kg/d DM, 68 g/d dietary N, and 70 g/d dietary starch. Starch flow to the duodenum was small (38 g/d), and N flow was 91 g/d. Small intestinal digestibility of duodenal N was 57%, and small intestinal digestion of duodenal starch flow was extensive (92%). Small intestinal starch digestibility was 34% when 1.5 kg/d raw cornstarch was continuously infused into the duodenum. Subsequently, cattle were placed in 1 of 2 replicated Latin squares that were balanced for carryover effects to determine response to casein infusions and time required for adaptation. Duodenal infusion of casein linearly increased (P ≤ 0.05) small intestinal starch digestibility, and small intestinal starch digestion adapted to infusion of casein in 6 d. Ethanol-soluble starch and unpolymerized glucose flowing to the ileum increased linearly (P ≤ 0.05) with increasing infusion of casein. Plasma cholecystokinin was not affected by casein infusion, but circulating levels of glucose were increased by casein supplementation (P ≤ 0.05). Responses in small intestinal starch digestion in cattle adapted to casein within 6 d, and increases in duodenal supply of casein up to 400 g/d increased small intestinal starch digestion in cattle.

Described is a method for producing copolymerized sol-gel derived sorbent particles for the production of copolymerized sol-gel derived sorbent material. The method for producing copolymerized sol-gel derived sorbent particles comprises adding a basic solution to an aqueous metal alkoxide mixture for a pH.ltoreq.8 to hydrolyze the metal alkoxides. Then, allowing the mixture to react at room temperature for a precalculated period of time for the mixture to undergo an increased in viscosity to obtain a desired pore size and surface area. The copolymerized mixture is then added to an immiscible, nonpolar solvent that has been heated to a sufficient temperature wherein the copolymerized mixture forms a solid upon the addition. The solid is recovered from the mixture, and is ready for use in an active sampling trap or activated for use in a passive sampling trap.

This invention relates to an electrophoresis separation medium having a gel matrix of at least one random, linear copolymer comprising a primary comonomer and at least one secondary comonomer, wherein the comonomers are randomly distributed along the copolymer chain. The primary comonomer is an acrylamide or an acrylamide derivative that provides the primary physical, chemical, and sieving properties of the gel matrix. The at least one secondary comonomer imparts an inherent physical, chemical, or sieving property to the copolymer chain. The primary and secondary comonomers are present in a ratio sufficient to induce desired properties that optimize electrophoresis performance. The invention also relates to a method of separating a mixture of biological molecules using this gel matrix, a method of preparing the novel electrophoresis separation medium, and a capillary tube filled with the electrophoresis separation medium.

Drying technology of angiotensin converting enzyme (ACE) inhibitory peptides derived from bovine casein was investigated. No significance was observed on ACE inhibitory activity of products prepared by spay drying and freeze drying (P>0.05). Spay drying was the best drying process for practical industry production. The inlet temperature ranged from 140℃ to 160℃ and the exit temperature ranged from 70℃ to 90℃ during the spay drying process. Under the optimal eonditious, scale-up of angiotensin converted enzyme inhibitory peptide from 1 L to 10 L and the experiment was successively conducted. Peptide yield was 29% and half inhibitory concentration(IC50) was 0.53g·L-1.

Casein variants occurring in milks from goats homozygous at the alpha(s1)-Cn locus were separated and identified by an RP-HPLC/ESI-MS method. Preferential haplotypes arose as well as some particularities in posttranslational modifications. A new variant of caprine beta-Cn, named C, as well as the phosphorylations pattern of the protein were characterized by the combined use of peptide mass fingerprinting and sequencing by tandem mass spectrometry. The molecular mass of the new variant in its 6P form was measured as 23854 Da and it differs in a mono amino acid substitution, A177 --> V177, from the variant A. The phosphorylation pattern of caprine beta-Cn is homologous to bovine beta-Cn concerning the 5P located on Ser15, 17, 18, 19, 35 but it presents a specific additional phosphorylation site on Thr12 that is comparable to human beta-Cn phosphorylation located on Thr3.

Full Text Available Background: Caries process is characterized by the presence of demineralization. Demineralization is caused by organic acids as a result of carbohydrate substrate fermentation. Remineralization is a natural repair process for non-cavitated lesions. Remineralization occurs if there are Ca2+ and PO43- ions in sufficient quantities. Casein-amorphous calcium phosphate phosphopeptide (CPP-ACP is a paste material containing milk protein (casein, that actually contains minerals, such as calcium and phosphate. The casein ability to stabilize calcium phosphate and enhance mineral solubility and bioavailability confers upon CPP potential to be biological delivery vehicles for calcium and phosphate. Purpose: The aim of this study was to determine the calcium levels in tooth enamel after being covered with CPP-ACP 2 times a day for 3, 14 and 28 days. Methods: Sample were bovine incisors of 3 year old cows divided into 4 groups, namely group I as control group, group II, III and IV as treatment groups covered with CPP-ACP 2 times a day. All of those teeth were then immersed in artificial saliva. Group II was immersed for 3 days, while group III was immersed for 14 days, and group IV was immersed for 28 days. One drop of CPP-ACP was used to cover the entire labial surface of teeth. The measurement of the calcium levels was then conducted by using titration method. All data were analyzed by One- Way ANOVA test with 5% degree of confidence. Results: The results showed significant difference of the calcium levels in tooth enamel of those groups after covered with CPP-ACP 2 times a day for 3, 14 and 28 days (p = 0.001. There is also significant difference of the calcium levels in tooth enamel of those treatment groups and the control group (p = 0.001. Conclusion: The calcium levels of tooth enamel are increased after covered with CPP-ACP 2 times a day for 3, 14 and 28 days.Latar belakang: Proses terjadinya karies gigi ditandai oleh adanya demineralisasi

Casein kinase 1δ (CK1δ) and 1ε (CK1ε) are believed to be necessary enzymes for the regulation of circadian rhythms in all mammals. On the basis of our previously published work demonstrating a CK1ε-preferring compound to be an ineffective circadian clock modulator, we have synthesized a series of pyrazole-substitued pyridine inhibitors, selective for the CK1δ isoform. Additionally, using structure-based drug design, we have been able to exploit differences in the hinge region between CK1δ and p38 to find selective inhibitors that have minimal p38 activity. The SAR, brain exposure, and the effect of these inhibitors on mouse circadian rhythms are described. The in vivo evaluation of these inhibitors demonstrates that selective inhibition of CK1δ at sufficient central exposure levels is capable of modulating circadian rhythms.

The enzymatic hydrolysates of casein are so complex that there is no effective method to do quantitative analysis.The common techniques,such as high performance chromatography and SDS-PAGE,can only carry out qualitative analysis.On the basis of membrane separation and high performance size exclusion chromatography (HPSEC),standard peptides with different molecular mass range were prepared,and the linear relationships between mass concentration of the standard peptides and the ultraviolet absorption of corresponding peak areas were established.Consequently,mass concentration of the different hydrolysates at different reaction times could be accurately calculated.The combination of chromatography and membrane separation is of great importance to the quantitative analysis of the complex hydrolysates,which can also be applied to the other macromolecular systems,such as carbohydrates.

The prevalence of classic autism and autism spectrum disorder (ASD) appears to be on the rise, and to date, there remains no clear etiology or cure. Out of desperation, many families are turning to new therapies and interventions discovered through various media sources and anecdotal reports from other parents. Unfortunately, many of these newer, well-publicized interventions have little empirical support. One of the most popular yet currently scientifically unproven interventions for ASD is the gluten-free, casein-free (GFCF) diet. Clinicians working with families of individuals with ASD are often asked for advice and find themselves unable to offer the most up-to-date and scientifically credible information. This article provides an overview of ASD and the GFCF diet, a summary and critique of current research findings, recommendations for future research, and practical advice for families to use in deciding if a trial of the GFCF diet is in the best interest of their child and family.

cDNA encoding the casein kinase II (CKII) subunits alpha and beta of human origin were expressed in Escherichia coli using expression vector pT7-7. Significant expression was obtained with E. coli BL21(DE3). The CKII subunits accounted for approximately 30% of the bacterial protein; however, most...... of the expressed proteins were produced in an insoluble form. The recombinant CKII alpha subunit was purified by DEAE-cellulose chromatography, followed by phosphocellulose and heparin-agarose chromatography. The recombinant CKII beta subunit was extracted from the insoluble pellet and purified in a single step...... on phosphocellulose. From 10 g bacterial cells, the yield of soluble protein was 12 mg alpha subunit and 5 mg beta subunit. SDS/PAGE analysis of the purified recombinant proteins indicated molecular masses of 42 kDa and 26 kDa for the alpha and beta subunits, respectively, in agreement with the molecular masses...

Full Text Available Beta-lactoglobulin (b-Lg and kappa-casein (k-Cn are two of the most importantproteins in the mammals’ milk synthesized by the epithelial cells of the mammaryglands. They play a crucial role in the milk quality and coagulation process(production of cheese and butter. The PCR-RFLP test was performed to distinguishthe different alleles in a population of Romanian Black Spotted cattle, a dairy breed.Genetic polymorphism was detected by digestion with the endonucleases Hae III (b-Lg and Hinf I (k-Cn, followed by electrophoresis in agarose high resolution gelstained with ethidium bromide. Fifty DNA samples from Romanian Black Spottedbreed were analyzed for A and B variants. This simple PCR-RFLP test makesfeasible the inclusion of b-Lg and k-Cn genotypes in breeding plans and cattleselection.

In mammals, the mechanism for the generation of circadian rhythms and entrainment by light–dark (LD) cycles resides in the hypothalamic suprachiasmatic nuclei (SCN), and the principal signal that adjusts this biological clock with environmental timing is the light:dark cycle. Within the SCN, rhythms are generated by a complex of molecular feedback loops that regulate the transcription of clock genes, including per and cry. Posttranslational modification plays an essential role in the regulation of biological rhythms; in particular, clock gene phosphorylation by casein kinase I, both epsilon (CKI) and delta (CKI), regulates key molecular mechanisms in the circadian clock. In this paper, we report for the first time that CKI activity undergoes a significant circadian rhythm in the SCN (peaking at circadian time 12, the start of the subjective night), and its pharmacological inhibition alters photic entrainment of the clock, indicating that CKI may be a key element in this pathway.

Full Text Available K-casein (K-CN represents one of the most important proteins determining the manufacturing properties of milk,because of its essential role in micelle formation and stabilisation.Several genetic variants of K-CN have been described in goats. To investigate the occurrence of seven alleles and theirdistribution among breeds, a total of 170 animals, from six different breeds reared in Italy (Cilentana Nera, Derivata diSiria, Maltese, Jonica, Garganica and Cashmere, have been analysed in this paper by the primer extension method.Alleles A and B were found to be the most represented in all the analysed breeds; allele D is present only in Maltese andCashmere animals with a very low frequency; while allele G has been found in all but two (Garganica and Cashmerebreeds. Alleles C, E and F were not present in the material used for this study.

In the present investigation, the preparation and characterization of a curcumin nanoemulsion with milk protein (sodium caseinate) and its incorporation into ice cream were undertaken. Among the different combinations, the most stable formulation was observed using milk fat (8%), medium chain triglycerides (2%), curcumin (0.24%) and sodium caseinate (6%) with a mean particle size of 333.8 ± 7.18 nm, a zeta potential of -44.1 ± 0.72 mV and an encapsulation efficiency of 96.9 ± 0.28%. The effect of different processing conditions (heating, pH and ionic strength) on the particle size distribution and zeta potential of the nanoemulsion was evaluated. During heat treatment, the particle size of the nanoemulsion was increased from 333.8 ± 7.18 to 351.1 ± 4.04 nm. The nanoemulsion was destabilized at pH 4.6 and the particle size increased above and below pH 5.0. However, there was a slight increase in the particle size with a change in the ionic concentration. The release kinetics data suggested that in simulated gastro-intestinal digestion, the nanoemulsion was stable against pepsin digestion (a 5.25% release of curcumin), while pancreatic action led to a 16.12% release of curcumin from the nanoemulsion. Finally, our formulation was successfully incorporated into ice cream and the sensory attributes were evaluated. No significant difference was observed in the scores of the sensory attributes between the control and ice cream prepared with a curcumin nanoemulsion. Moreover, the encapsulation efficiency of the curcumin incorporated into the ice cream was 93.7%, which indicates that it can withstand the processing conditions. The findings suggest that ice cream is a suitable dairy product for the delivery of lipophilic bioactive components (curcumin) which can be used for therapeutic purposes.

Full Text Available BACKGROUND: The generation of antibodies is impaired in newborns due to an immature immune system and reduced exposure to pathogens due to maternally derived antibodies and placental functions. During nursing, the immune system of newborns is challenged with multiple milk-derived proteins. Amongst them, caseins are the main constituent. In particular, human αS1-casein (CSN1S1 was recently shown to possess immunomodulatory properties. We were thus interested to determine if auto-antibodies to CSN1S1 are induced by breast-feeding and may be sustained into adulthood. METHODS: 62 sera of healthy adult individuals who were (n = 37 or were not (n = 25 breast-fed against human CSN1S1 were investigated by a new SD (surface display-ELISA. For cross-checking, these sera were tested for anti Epstein-Barr virus (EBV antibodies by a commercial ELISA. RESULTS: IgG-antibodies were predominantly detected in individuals who had been nursed. At a cut-off value of 0.4, the SD-ELISA identified individuals with a history of having been breast-fed with a sensitivity of 80% and a specificity of 92%. Under these conditions, 35 out of 37 sera from healthy donors, who where breast-fed, reacted positively but only 5 sera of the 25 donors who were not breast-fed. The duration of breast-feeding was of no consequence to the antibody reaction as some healthy donors were only short term breast-fed (5 days minimum until 6 weeks maximum, but exhibited significant serum reaction against human CSN1S1 nonetheless. CONCLUSION: We postulate that human CSN1S1 is an autoantigen. The antigenicity is orally determined, caused by breast-feeding, and sustained into adulthood.

Charge screening effects on nanostructures of N-isopropylacrylamide-sodium acrylate (NIPA-SA) and -acrylic acid (NIPA-AAc) gels are investigated with small-angle neutron scattering. The NIPA-SA and NIPA-AAc gels with low water content exhibit microphase separations with different dimensions....... The dehydrated NIPA-SA gel also makes the microphase separation but the dehydrated NIPA-AAc gel does not. These results indicate that ionic circumstance around charged bases strongly affects the nanostructures both of the dehydrated gel and the gel with low water content. (C) 2004 Elsevier B. V. All rights...

Syneresis of chitin gels formed in the course of N-acetylation of chitosan in hydroalcoholic media has been studied. A critical cross-linking density related to a critical acetylation degree for which the gel undergoes weak syneresis and swells in water was shown (degree of acetylation (DA) 88%). Above this value, the weight loss during syneresis increases with DA. Conversely, syneresis decreases on increasing the polymer concentration, but disappears at a macroscopic level for a polymer concentration close to the critical concentration of entanglement in the initial solution. An increase in temperature favours the formation of hydrophobic interactions and new inter- and intramolecular hydrogen bondings. Due to the weak polyelectrolyte character of chitin, the weight of the gel depends on the pH and ionic strength of the media. Swelling-deswelling experiments show that the swelling of the gel is not fully reversible in relation with the formation of new cross-links during the depletion of the network. Our results reveals that the balance between segment-segment and segment-solvent interactions as well as the molecular mobility play the major role.

AIM: The aim of this conference paper was to systematically review the quality of evidence and summarize the findings of clinical trials published after 2002 using fluoride mouth rinses, fluoride gels or foams for the prevention of dental caries. METHODS: Relevant papers were selected after an el...... brushing with fluoride toothpaste....

Alginate is a naturally occurring anionic polysaccharide extracted from brown algae. Because of biocompatibility, low toxicity, and simple gelation process, alginate gels are used in biomedical and food applications. Here, we report the rheological behavior of ionically crosslinked alginate gels, which are obtained by in situ gelation of alginates with calcium salts, in between two parallel plates of a rheometer. Strain stiffening behavior was captured using large amplitude oscillatory shear (LAOS) experiments. In addition, negative normal stress was observed for these gels, which has not been reported earlier for any polysaccharide networks. The magnitude of negative normal stress increases with applied strain and can exceed that of the shear stress at large strain. Rheological results fitted with a constitutive model that considers both stretching and bending of chains indicate that nonlinearity is likely related to the stretching of the chains between the crosslink junctions. The results provide an improved understanding of the deformation mechanism of ionically crosslinked alginate gel and the results will be important in developing synthetic extracellular matrix (ECM) from these materials.

Colostrum is a complex mixture of bioactives that promotes neonate growth. Recently, we have found by in vivo study that skimmed, sterilized, and concentrated bovine late colostrum (SCBLC), obtained from a Holstein herd on days 6-7 after parturition, had an ability to maintain intestinal integrity. In the present study we investigated effects of SCBLC on rat intestinal IEC-6 cell proliferation in vitro. A fraction containing αs1-casein was found to have a robust stimulation effect as compared to other protein fractions from SCBLC and even the αs1-casein fraction from milk from other Holstein herds. Furthermore, the SCBLC αs1-casein molecule demonstrated not only slightly slower mobility on both SDS- and native-PAGE than other bovine milk αs1-caseins, but also a peculiar conformation reminiscent of moltenglobule in the circular dichroism spectrum. These findings may be of relevant to the competence of SCBLC to preserve intestinal integrity.

Full Text Available In cattle, sheep and goats (Hayes et al., 1993 the casein genes form a cluster, on the chromosome 6, of four tightly linked genes, whose sequence is: αS1-β-αS2-k (Threadgill and Womack, 1990; Ferretti et al., 1990; Gallagher et al., 1994; Rijnkles et al., 1997. For this reason the alleles at these loci are generally inherited all together (Di Stasio and Mariani, 2000, with low recombination frequency. Thus it is worth noting the importance of describing the casein haplotypes namely the inherited combination of closely linked alleles (King and Stansfield, 1985; Meuwissen and Goddard, 2001. The knowledge of the casein allele associations could be important for the selection. In goat milk the casein alleles are related to important differences in the level of synthesis of the respective gene products and in milk chemical-physical properties........

to a single bolus intake of whey or casein after performance of heavy resistance exercise. Young male individuals were randomly assigned to participate in two protein trials (n = 9) or one control trial (n = 8). Infusion of l-[1-(13)C]leucine was carried out, and either whey, casein (0.3 g/kg lean body mass......), or a noncaloric control drink was ingested immediately after exercise. l-[1-(13)C]leucine-labeled whey and casein were used while muscle protein synthesis (MPS) was assessed. Blood and muscle tissue samples were collected to measure systemic hormone and amino acid concentrations, tracer enrichments......, and myofibrillar protein synthesis. Western blots were used to investigate the Akt signaling pathway. Plasma insulin and branched-chain amino acid concentrations increased to a greater extent after ingestion of whey compared with casein. Myofibrillar protein synthesis was equally increased 1-6 h postexercise after...

Here, we identified the milk protein α-casein as a novel suppressor of tumor growth and metastasis. Briefly, Met-1 mammary tumor cells expressing α-casein showed a ~5-fold reduction in tumor growth and a near 10-fold decrease in experimental metastasis. To identify the molecular mechanism(s), we performed genome-wide transcriptional profiling. Interestingly, our results show that α-casein upregulates gene transcripts associated with interferon/STAT1 signaling and downregulates genes associated with "stemness." These findings were validated by immunoblot and FACS analysis, which showed the upregulation and hyperactivation of STAT1 and a decrease in the number of CD44(+) "cancer stem cells." These gene signatures were also able to predict clinical outcome in human breast cancer patients. Thus, we conclude that a lactation-based therapeutic strategy using recombinant α-casein would provide a more natural and non-toxic approach to the development of novel anticancer therapies.

Casein is the main component of milk protein,accounts for about 70~80 percent of total protein.as1-casein is the main component of casein,accounting for about 39~46 percent of total milk protein.It is one of the quite active functional genes in the cattle＇s mammary gland tissue,so it is important and very necessary to study the cow alpha S1-casein gene.The paper summarized the gene structure,genetic diversity and casein active peptide of alpha-s1-casein gene,which would provide the theory basis to study alpha-s1-casein gene deeply and to improve the milk protein content by regulating as1-casein gene.%酪蛋白是牛奶蛋白质的主要组成部分,约占总蛋白的70%~80%,而as1-酪蛋白又是酪蛋白的主要组成部分,约占牛奶总蛋白的39%~46%,是牛乳腺组织中转录相当活跃的一个功能基因,因此研究牛αS1-酪蛋白基因就显得尤为重要而且很有必要。本文从牛as1-酪蛋白基因结构、遗传多态性和酪蛋白活性肽方面做了综述,旨在为以后牛as1-酪蛋白基因的深入研究和通过调控as1-酪蛋白基因而提高牛奶蛋白质的含量提供理论基础。

Using Saanen ＆ Guanzhong goat milk as material,caprine casein was obtained from the milk. The effects of pH,temperature,ionic calcium,citrate,phosphate and casein on the heat stability of caprine casein were studied by hot coagulation time （HCT）. The results showed that the heat stability of casein was better at the pH of 6.8, high temperature can lower the heat stability,ionic calcium can speed up the HCT of casein. A certain amount of citrate or phosphate can effectively raise the hot stability of caprine casein. The addition of casein was not obvious to the heat stability of caprine casein.%以莎能和关中羊奶为原料，通过从羊奶中提取酪蛋白，分别在不同的pH、温度以及添加不同浓度的Ca^2＋、柠檬酸钠、三聚磷酸钠、干酪素的条件下测定酪蛋白的热凝固时间（HCT），研究其对羊奶酪蛋白热稳定性的影响。结果表明，pH在6．8时酪蛋白的热稳定性最好，高温会降低酪蛋白的热稳定性，钙离子可以降低羊奶酪蛋白的热稳定性，适量的柠檬酸钠或三聚磷酸钠可以有效提高羊奶酪蛋白的热稳定性，干酪素对酪蛋白稳定性影响不明显。

Full Text Available The aim of this study was to introduce a reference method for the detection of cow milk in ewe and goat cheeses (EC 273/08 in order to protect manufacturers and consumers from adulterations and imitations, and to ensure alignment with the demands of domestic and EU markets. The method includes isolation of casein from cheese, isoelectric focusing of γ2- and γ3-casein originating from the hydrolysis of β-casein by plasmin, the detection and quantitative determination of γ-casein in cow, ewe and goat cheese by densitometry. Ewe or goat cheese products with a minimum of 1 % of cow milk are considered to be adulterated. For the quantitative determination of cow, ewe and goat milk in cheeses, standard mixtures of cow-ewe and cow-goat milk were made by adding 0; 0.5; 1; 2; 5; 10; 25; 50; 75 and 100 % (v/v of cow milk. The quantification was performed by determining the peak area ratio of cow γ-casein in comparison to ewe/goat casein in prepared standard cheeses. The calibration curves were calculated based on the relation of the peak area ratio of cow γ-caseins (calculated as the percentage of total γ-caseins in contrast to the relative content (% of cow milk in the mixture. The method proved to be adequate for the detection of raw and heat-treated cow milk in fresh and ripened cheeses made from ewe or goat milk, or a mixture of ewe and goat milk.

International audience; Protein ingredients such as hydrolysates of milk proteins may improve the nutritive value of functional foods. For instance, $\\beta$-casein tryptic hydrolysate could simultaneously increase iron absorption and prevent lipid oxidation in foods containing high contents of polyunsaturated fatty acids (PUFA). The aim of this study was to determine the antioxidant activity of $\\beta$-casein tryptic hydrolysate and of its $\\beta$-caseinophosphopeptide (f1-25) on Fe(III)/asco...

Pluronic F127 solutions form gels in water with high elastic moduli. Pluronic gels can, however, only withstand small deformations and stresses. Different steady shear and oscillatory methods traditionally used to determine yield stress values are compared. The results show that the yield stresses...... values of these gels depend on test type and measurement time, and no absolute yield stress value can be determined for these physical gels....

@@ 1 Introduction Polymer gels have been extensively studied[1～17] since the discovery of volume phase-transition of a gel by Tanaka[1～5]. As a unique soft material, gels attract much attention and are tried to be applied for drug-delivery systgems[6], actuators or chemo-mechanical devices[7～9] and so on. In particular, controlled-release of small molecules from a gel is now a subject of special interest[10].

The Fricke xylenol gel (FXG) dosimeter has been studied for application in radiotherapy because it is capable of to measure the spatial distribution of radiation doses. The dosimetry is based on the oxidation of ferrous (Fe2+) to ferric (Fe3+) ions radiation induced, related to the radiation dose. The gel material usually employed is the 300 Bloom gelatin, which is imported and very expensive in Brazil. Aiming to analyze the viability of to use a locally produced and low cost gel material, in this work the spectrophotometric responses of FXG solutions prepared using 270 Bloom gelatin commercially available and 300 Bloom gelatin imported were compared. The absorption spectra of solutions prepared with 5% by weight 270 and 300 Bloom gelatins non-irradiated and irradiated with 60Co gamma radiation in the dose range between 0.5 and 100 Gy were analysed, the dose-response curves were evaluated and the useful dose range was established. The obtained results indicate that the FXG solution prepared with 270 Bloom gelatin presents good performance, similar to that presented by the FXG solution prepared with 300 Bloom gelatin and its use can be recommended owing to the low cost and the availability in local market.

Detailed characterization of phosphoproteins as well as other post-translationally modified proteins is required to fully understand protein function and regulatory events in cells and organisms. Here we present a mass spectrometry (MS) based experimental strategy for the identification and mapping...... chromatography (Fe(III)-IMAC) columns were employed for enrichment of phosphorylated peptides from crude peptide mixtures prior to off-line analysis by matrix-assisted laser desorption/ionization (MALDI) MS or nanoelectrospray tandem mass spectrometry (MS/MS). An optimized and sensitive procedure for alkaline...... of phosphorylation sites. The advantages and limitations of the experimental strategy was demonstrated by enrichment, identification and sequencing of phosphopeptides from the model proteins ovalbumin and bovine beta-casein isolated by gel electrophoresis. Furthermore, an autophosphorylation site at Ser-3...

β-Carotene is a carotenoid usually applied in the food industry as a precursor of vitamin A or as a colourant. β-Carotene is a labile compound easily degraded by light, heat and oxygen. Casein micelles were used as nanostructures to encapsulate, stabilise and protect β-carotene from degradation during processing in the food industry. Self-assembly method was applied to re-assemble nanomicelles containing β-carotene. The protective effect of the nanostructures against degradation during the most common industrial treatments (sterilisation, pasteurisation, high hydrostatic pressure and baking) was proven. Casein micelles protected β-carotene from degradation during heat stabilisation, high pressure processing and the processes most commonly used in the food industry including baking. This opens new possibilities for introducing thermolabile ingredients in bakery products.

Full Text Available AlphaS1 casein fraction in caprine milk is characterized by an important polymorphism due to substitution, deletion of amino acids and post trascriptional modifications (Grosclaude et al., 1994; Ferranti et al., 1997. This structural polymorphism is associated to a quantitative variability in protein expression related to different milk quality and dairy properties (Pierre et al., 1998; Remeuf, 1993; Vassal et al., 1994. Classical electrophoretic methods were applied to characterize the phenotypic variants at αS1-casein fraction (Addeo et al., 1988; Russo et al., 1986. During the last ten years capillary electrophoresis became an analytical technique for rapid and automated analysis requiring small sample volume and small solvent waste. These characteristics, together with the high resolution and the chance to give quantitative results, made this technique a useful tool........

Because of variable degrees of phosphorylation, alternative splicing, and probable instability resulting from nonenzymatic deamidation, equine beta-casein presents a complex pattern by 2-dimensional electrophoresis that needs clarification. beta-Casein prepared from Haflinger mare's milk by hydrophobic interaction chromatography was fractionated by ion-exchange chromatography according to the degree of phosphorylation. Isoforms were identified by mass spectrometry; they corresponded to the full-length protein having 3 to 7 phosphate groups and to the splicing variant involving exon 5 and containing 4 to 7 phosphate groups. Investigations of nonenzymatic deamidation showed that beta-casein did not deamidate spontaneously in stored milk and during the different steps of chromatography, but deamidation could occur when 2-dimensional electrophoresis was performed, increasing the beta-casein pattern complexity. This phenomenon was strongly minimized when the first dimension step was carried out at 10 degrees C instead of at room temperature. Finally, spot attribution on 2-dimensional pattern of beta-casein was achieved by mixing each phosphorylation isoform in its native state with the whole beta-casein fraction.

To better understand sensory perception of foods, water exudation studies on protein-based gels are of a high importance. It was aimed to study the interplay of gel coarseness and gel stiffness on water holding (WH) and water flow kinetics from the gel once force is applied onto the material. Ovalbu

1 Results It has been reported that crystalline form racemates can be resoluted by various methods[1],but for optical resolution of liquid form racemates,the method is limited in chromatograph.So chiral compounds cannot be obtained on large scale by such method.In our previous paper,(+)-3-chloro-1,2-propanediol has been obtained by optical resolution of its racemata (liquid) with a chiral biopolymer,casein[2].

Full Text Available The effects of daily dietary Bacillus subtilis (Bs, and adding L-tryptophan, fructan, or casein to fecal fermentation broths were investigated as means to reduce the production of noxious gas during manure fermentation caused by ammonia, hydrogen sulfide (H2S, and 3-methylindole (skatole. Eighty swine (50.0±0.5 kg were equally apportioned to an experimental group given Bs in daily feed, or a control group without Bs. After 6 weeks, fresh manure was collected from both groups for fermentation studies using a 3×3 orthogonal array, in which tryptophan, casein, and fructan were added at various concentrations. After fermentation, the ammonia, H2S, L-tryptophan, skatole, and microflora were measured. In both groups, L-tryptophan was the principle additive increasing skatole production, with significant correlation (r = 0.9992. L-tryptophan had no effect on the production of ammonia, H2S, or skatole in animals fed Bs. In both groups, fructan was the principle additive that reduced H2S production (r = 0.9981. Fructan and Bs significantly interacted in H2S production (p = 0.014. Casein was the principle additive affecting the concentration of ammonia, only in the control group. Casein and Bs significantly interacted in ammonia production (p = 0.039. The predominant bacteria were Bacillus spp. CWBI B1434 (26% in the control group, and Streptococcus alactolyticus AF201899 (36% in the experimental group. In summary, daily dietary Bs reduced ammonia production during fecal fermentation. Lessening L-tryptophan and increasing fructan in the fermentation broth reduced skatole and H2S.

To obtain information on the safety and efficacy of the gluten-free/casein-free (GFCF) diet, we placed 14 children with autism, age 3-5 years, on the diet for 4-6 weeks and then conducted a double-blind, placebo-controlled challenge study for 12 weeks while continuing the diet, with a 12-week follow-up. Dietary challenges were delivered via weekly…

Prolactin receptors were partially purified from rabbit mammary gland membranes by using an affinity chromatography technique. Antibodies against this prolactin receptor preparation were obtained in guinea pig and sheep. Both antisera were able to inhibit the binding of 125I-labeled ovine prolactin to rabbit mammary gland membranes. When added to culture media of rabbit mammary explants, the anti-prolactin receptor antiserum inhibited the capacity of prolactin to initiate casein synthesis and...

Full Text Available Abstract Background Postprandial metabolic and appetitive responses of proteins are dependent on protein source and processing technique prior to ingestion. Studies on the postprandial effects of enzymatic crosslinking of milk proteins are sparse. Our aim was to study the effect of transglutaminase (TG-induced crosslinking of sodium caseinate on postprandial metabolic and appetite responses. Whey protein was included as reference protein. Methods Thirteen healthy individuals (23.3 ± 1.1 y, BMI 21.7 ± 0.4 kg/m2 participated in a single-blind crossover design experiment in which the subjects consumed three different isovolumic (500 g pourable beverages containing either sodium caseinate (Cas, 29 g, TG-treated sodium caseinate (Cas-TG, 29 g or whey protein (Wh, 30 g in a randomized order. Blood samples were collected at baseline and for 4 h postprandially for the determination of plasma glucose, insulin and amino acid (AA concentrations. Gastric emptying (GE was measured using the 13 C-breath test method. Appetite was assessed using visual analogue scales. Results All examined postprandial responses were comparable with Cas and Cas-TG. The protein type used in the beverages was reflected as differences in plasma AA concentrations between Wh and Cas, but there were no differences in plasma glucose or insulin responses. A tendency for faster GE rate after Wh was detected. Appetite ratings or subsequent energy intake did not differ among the protein beverages. Conclusions Our results indicate that the metabolic responses of enzymatically crosslinked and native sodium caseinate in a liquid matrix are comparable, suggesting similar digestion and absorption rates and first pass metabolism despite the structural modification of Cas-TG.

Full Text Available Abstract Background & aim: Due to the low solubility and high toxicity of drugs, treatment of cancers is problematic therefore, the encapsulation and targeted delivery of therapeutic effect is required. The aim of this study was to investigate the effect of nanoparticles on cellular uptake and bioavailability of beta-casein on platinum complexes as cancer drugs. Methods: In the present experimental study, the physicochemical properties of nanoparticles as drug carriers of beta-casein devices using dynamic light scattering (DLS and scanning electron microscopy (SEM were investigated. In order to evaluate the toxicity effects of platinum complexes, the colon cancer cells in the absence or presence of free platinum complex concentration and nanoparticle loaded with platinum complexes were incubated for 24 and 48 hours. LD50 Values (concentration of compound causing 50% mortality in the cells was determined using the MTT assay. Data were analyzed by ANOVA and post-hoc test. Results: At a concentration of 1 mg ml, beta-casein nanoparticle drug carriers were synthesized in the range of 100 to 300 µM. In addition, the mortality rate in cancer cells by the release of platinum complexes (without and with the capsule, were 70 and 26 in 24 hours, and 60 µM and 21 µM in 48 hours respectively, Conclusion: The study showed that the bioavailability of the encapsulated platinum complexes increases and new drug delivery system may be a good candidate for the treatment of cancer. Key words: Beta-casein, Pt (II Complex, Bioavalibility, Nanocarrier, Micelle

Most current research has focused on using ceramic microfiltration (MF) membranes for micellar casein concentrate production, but little research has focused on the use of polymeric spiral-wound (SW) MF membranes. A method for the production of a serum protein (SP)-reduced micellar casein concentrate using SW MF was compared with a ceramic MF membrane. Pasteurized (79°C, 18s) skim milk (1,100 kg) was microfiltered at 50°C [about 3 × concentration] using a 0.3-μm polyvinylidene fluoride spiral-wound membrane, bleed-and-feed, 3-stage process, using 2 diafiltration stages, where the retentate was diluted 1:2 with reverse osmosis water. Skim milk, permeate, and retentate were analyzed for SP content, and the reduction of SP from skim milk was determined. Theoretically, 68% of the SP content of skim milk can be removed using a single-stage 3× MF. If 2 subsequent water diafiltration stages are used, an additional 22% and 7% of the SP can be removed, respectively, giving a total SP removal of 97%. Removal of SP greater than 95% has been achieved using a 0.1-μm pore size ceramic uniform transmembrane pressure (UTP) MF membrane after a 3-stage MF with diafiltration process. One stage of MF plus 2 stages of diafiltration of 50°C skim milk using a polyvinylidene fluoride polymeric SW 0.3-μm membrane yielded a total SP reduction of only 70.3% (stages 1, 2, and 3: 38.6, 20.8, and 10.9%, respectively). The SP removal rate for the polymeric SW MF membrane was lower in all 3 stages of processing (stages 1, 2, and 3: 0.05, 0.04, and 0.03 kg/m(2) per hour, respectively) than that of the comparable ceramic UTP MF membrane (stages 1, 2, and 3: 0.30, 0.11, and 0.06 kg/m(2) per hour, respectively), indicating that SW MF is less efficient at removing SP from 50°C skim milk than the ceramic UTP system. To estimate the number of steps required for the SW system to reach 95% SP removal, the third-stage SP removal rate (27.4% of the starting material SP content) was used to

Full Text Available Background: Autism is a complex psychiatric disorder characterized by three core symptoms, i.e. impairments in social interaction, restricted patterns of behavior and impairments in communication. In the framework of the “opioid excess theory”, the disorder symptoms are compared to the behavioral effects of opiates. Based on this, a possible nutritional basis of autism has been proposed, hypothesizing that certain food proteins such as gluten and casein can be transformed to opioid peptides during digestion. These peptides might eventually be able to enter the blood stream and act upon the central nervous system. As a consequence, a diet low in such proteins has been hypothesized to ameliorate the behavioral symptoms of autistic children. Objective: The scope of this review was to analyze the effects of gluten-free and casein-free (GFCF diets on children with autism, as well as to provide information concerning additional aspects related to the GFCF diet in autism. Methods: A literature search was conducted including scientific publications up until December 2013. Search results were screened for any kind of GFCF dietary intervention as well as surveys dealing with GFCF as a treatment for autism. Results: A review of survey data shows that up to 25 % of parents of affected children report on current use of a GFCF diet. The majority of identified studies evaluating GFCF diet outcomes failed to meet basic methodological standards of interventional science. Comparison of study results did not show any clear-cut results, with a substantial proportion of studies failing to show any positive dietary effect. The results of more sophisticated trials were far from equivocal and the studies differed by many methodological aspects. Some variables such as information source and trial duration seemed to affect outcome. Conclusions: Evidence for the effectiveness of the GFCF diet in the treatment of autism is sparse. Rigorous scientific evaluations partly

Full Text Available Jennifer Harrison Elder,1 Consuelo Maun Kreider,2 Nancy M Schaefer,3 Mary B de Laosa4 1Department of Family and Community Health Nursing Science, 2Department of Occupational Therapy, 3Health Science Center Library, 4Department of Psychology, University of Florida, Gainesville, FL, USA Background: The gluten-free, casein-free (GFCF diet is heralded by strong anecdotal parental reports to greatly improve and even "cure" symptoms of autism spectrum disorders (ASDs. Yet, to date, little conclusive empirical evidence exists supporting its use. Objective: The purpose of this paper is to provide an overview of the state of the recent evidence regarding the use of GFCF diet for treatment of individuals with ASD. Methods: Five database providers (PubMed, Web of Knowledge, EBSCO, ProQuest, and WorldCat were used to search 19 databases, yielding a total of 491 articles that were published through February 2015. Peer-reviewed articles published between January 2005 and February 2015 were included for review if study participants were identified as having ASD and if the study investigated the effects of the GFCF diet on ASD behaviors or the relationship between the diet and these behaviors. Results: Evaluation of search results yielded eleven reviews, seven group experimental studies including five randomized controlled trials, five case reports, and four group observational studies published during the last 10 years. These studies represent a marked increase in the number of reported studies as well as increased scientific rigor in investigation of GFCF diets in ASD. Conclusion: While strong empirical support for the GFCF diet in ASD is currently lacking, studies point to the need for identifying subsets of individuals (eg, those with documented gastrointestinal abnormalities who may be the best responders to the GFCF diet. Identifying these subsets is critically needed to enhance rigor in this research area. Until rigorous research supporting the use of

AIM: The aim of this conference paper was to systematically review the quality of evidence and summarize the findings of clinical trials published after 2002 using fluoride mouth rinses, fluoride gels or foams for the prevention of dental caries. METHODS: Relevant papers were selected after...... an electronic search for literature published in English between 2003 and 2014. The included papers were assessed for their risk of bias and the results were narratively synthesized due to study heterogeneity. The quality of evidence was expressed according to GRADE. RESULTS: A total of 19 papers were included...... (6 on fluoride mouth rinse, 10 on fluoride gel and 3 on fluoride foam); 6 had a low risk of bias while 2 had a moderate risk. All fluoride measures appeared to be beneficial in preventing crown caries and reversing root caries, but the quality of evidence was graded as low for fluoride mouth rinse...

Gambarini, G. [Department of Physics of the University and INFN, Milan (Italy)]. e-mail: grazia.gambarini@mi.infn.it

2005-07-01

With the continuum development of conformal radio therapies, aimed at delivering high dose to tumor tissue and low dose to the healthy tissue around, the necessities has appeared of suitable improvement of dosimetry techniques giving the possibility of obtaining dose images to be compared with diagnostic images. Also if wide software has been developed for calculating dose distributions in the fields of various radiotherapy units, experimental verifications are necessary, in particular in the case of complex geometries in conformal radiotherapy. Gel dosimetry is a promising method for imaging the absorbed dose in tissue-equivalent phantoms, with the possibility of 3D reconstruction of the spatial dose distribution, with milli metric resolution. Optical imaging of gel dosimeters, based on visible light absorbance analysis, has shown to be a reliable technique for achieving dose distributions. (Author)

The synthesis of ceramic raw materials has become an important factor in ceramic technologies. The increasing demands to the performance of ceramic compounds has caused increased activities for the preparation of tailor-made raw materials. Amongst a variety of new syntheses like flame pyrolysis, reactive spray drying, plasma or laser assisted techniques, the sol-gel process plays an important and increasing role. The process describes the building up of an inorganic (in general an oxide) netw...

Abstract An increasingly important application of liquid jets is the disintegration of the jet to form droplets of liquid containing nuclear fuel. These droplets are then dried and sintered to form ceramic micro spheres for use in fuel elements in nuclear reactors. The total operations required to form the droplets convert them to solids and fire them to ceramic bodies comprise what are known as Sol-Gel processes Reference 13.

Casein genetic polymorphisms are important and well known due to their effects on quantitative traits and technological properties of milk. At the DNA level, polymerase chain reaction (PCR)-single-strand conformation polymorphism (SSCP) allows for the simultaneous typing of several alleles at casein loci, as well as the detection of unknown polymorphisms. Here we describe the usefulness of the PCR-SSCP technique for casein typing in sheep. In particular, three single-nucleotide polymorphisms (SNP) are described at CSN1S1, CSN2, and CSN3, all resulting in amino acid exchanges. At CSN1S1, a transition T-->C was found, resulting in the deduced amino acid exchange Ile186-->Thr186. A transition A-->G resulting in the deduced amino acid exchange Met183-->Val183 was identified at CSN2. The 2 SNP showed a rather high frequency (ranging from 0.12 to 0.26) in 3 Italian breeds (Sarda, Comisana, Sopravissana). Another transition C-->T (Ser104-->Leu104) was found at CSN3 in one heterozygous animal.

Full Text Available Caseins are a family of milk proteins that exist in several molecular forms and arethe main proteins present in the bovine milk. Genetic variation of these proteins hasbeen associated with the quality and quantity of cheese derived from milk.This study was focused on possibilities to evaluate the frequency of the K-casein Ballele in dairy herds from the Research and Development Station for Bovine RaisingArad in order to have breeding programs that target an increase in the frequency ofthe B allele in the dairy cattle population.In order to differentiate the favorable genotype for superior composition and highercheese yield, we used simple DNA extraction method from fresh blood andtechniques based on DNA analysis, which include polymerase chain reaction andrestriction fragment length polymorphisms (PCR-RFLP methods. Employing thesetechniques we were able to determine the k-casein genotype of all individuals in agiven population under selection, regardless of sex, age or physiological stage.As a result, it is now possible to include information on milk protein genotypes intomarker assisted selection programs and consequently improve response to selection.

In this study, structural changes in micellar caseins and whey proteins due to high pressure--low temperature treatments (HPLT) were investigated and compared to changes caused by high pressure treatments at room temperature. Whey protein isolate (WPI) solutions as well as micellar casein (MC) dispersions and mixtures were treated at 500 MPa (pH 7.0 and 5.8) at room temperature, -15 °C and -35 °C. Surface hydrophobicity and accessible thiol groups remained nearly unchanged after HPLT treatments whereas HP treatments at room temperature caused an unfolding of the WPI, resulting in an increase in surface hydrophobicity and exposure of the thiol groups. For HPLT treatments, distinct changes in the secondary structure (increase in the amount of β-sheets) were observed while the tertiary structure remained unchanged. Large flocs, stabilized by hydrophobic interactions and hydrogen bonds, were formed in casein containing samples due to HPLT treatments. Depending on the pH and the applied HPLT treatment parameters, these interactions differed significantly from the interactions determined in native micelles.

In this study peptic and tryptic digestions of bovine {beta}-casein were investigated using quartz crystal microbalance (QCM). {beta}-Casein, which was used as a model protein, was immobilized on the surface of the QCM sensor where its degradation caused shifts in the resonant frequency. Atomic force microscopy was applied for the characterization of the protein layer. Different pH-values for peptic or tryptic digestions were chosen to visualize their effect on enzyme activity. Lower frequency shifts were observed at pH-values deviating from those at the maximum enzyme activity. In the case of the peptic digestion the frequency shift at pH 4 was more than 10 times smaller than those at pH 2. The frequency shifts for tryptic digestions at pH 5.4 and pH 6.4 were about two thirds compared to that obtained for the digestion at pH 7.4. The identification of peptides using MALDI-ToF mass spectrometry was used for verification of the proteolyses of the immobilized protein. Furthermore, it was shown that the QCM technique allows close observation of the effect of different pH-values on the immobilized casein layer. All in all, QCM facilitates the monitoring of the progress of enzymatic reactions in real-time.

The objective of the present study was to investigate the effect of environmental factors, milk casein content and titratable acidity on milk coagulation properties (MCP) of samples routinely collected in the Trento province (northeast Italy) under field conditions. Rennet coagulation time (RCT, min), curd-firming time (k20, min) and curd firmness (a30, mm) were determined by Formagraph on 14 971 samples from 635 herds associated to 17 dairy factories. Besides MCP, fat, protein, and casein percentages, titratable acidity (TA), and somatic cell and bacterial counts were available. A standardised index of milk aptitude to coagulate (IAC) was derived using information of RCT and a30. An analysis of variance was conducted on MCP and IAC using a fixed effects linear model. Approximately 3% of milk samples did not form a curd within the testing time (30 min) and k20 was missing for 26% of milks. The percentage of samples without information on k20 largely differed among dairy factories (1·7-20·9%). Significant differences were estimated between the best and the worst dairy factory for RCT (-2 min), k20 (-1·2 min), a30 (+3·4 mm) and IAC (+2·6 points). Milk casein content and TA were important factors in explaining the variation of MCP and IAC, supporting the central role of these two traits on technological properties. The Trento province is heterogeneous in terms of dairy systems and this could explain the differences among dairy factories.

Full Text Available Caseins are a family of milk proteins that exist in several molecular forms and arethe main proteins present in the bovine milk. Genetic variation of these proteins hasbeen associated with the quality and quantity of cheese derived from milk.This study was focused on possibilities to evaluate the frequency of the K-casein Ballele in dairy herds from the Research and Development Station for Bovine RaisingArad in order to have breeding programs that target an increase in the frequency ofthe B allele in the dairy cattle population.In order to differentiate the favorable genotype for superior composition and highercheese yield, we used simple DNA extraction method from fresh blood andtechniques based on DNA analysis, which include polymerase chain reaction andrestriction fragment length polymorphisms (PCR-RFLP methods. Employing thesetechniques we were able to determine the k-casein genotype of all individuals in agiven population under selection, regardless of sex, age or physiological stage.As a result, it is now possible to include information on milk protein genotypes intomarker assisted selection programs and consequently improve response to selection.

The influence of milk protein-based ingredients on the textural characteristics, sensory properties, and microstructure of probiotic yogurt during a refrigerated storage period of 28 d was studied. Milk was fortified with 2% (wt/vol) skim milk powder as control, 2% (wt/vol) sodium calcium caseinate (SCaCN), 2% (wt/vol) whey protein concentrate (WPC) or a blend of 1% (wt/vol) SCaCN and 1% (wt/vol) WPC. A commercial yogurt starter culture and Bifidobacterium lactis Bb12 as probiotic bacteria were used for the production. The fortification with SCaCN improved the firmness and adhesiveness. Higher values of viscosity were also obtained in probiotic yogurts with SCaCN during storage. However, WPC enhanced water-holding capacity more than the caseinate. Addition of SCaCN resulted in a coarse, smooth, and more compact protein network; however, WPC gave finer and bunched structures in the scanning electron microscopy micrographs. The use of SCaCN decreased texture scores in probiotic yogurt; probably due to the lower water-holding capacity and higher syneresis values in the caseinate-added yogurt sample. Therefore, the textural characteristics of probiotic yogurts improved depending on the ingredient variety.

Understanding the mechanisms that determine the diffusion and interaction of macromolecules (such as proteins and polysaccharides) that disperse through dense media is an important fundamental issue in the development of innovative technological and medical applications. In the current work, the partitioning and diffusion of macromolecules of different sizes (from 4 to 10 nm in diameter) and shapes (linear or spherical) within dispersions of casein micelles (a protein microgel) is studied. The coefficients for diffusion and partition are measured using FRAP (fluorescence recovery after photobleaching) and analyzed with respect to the structural characteristics of the microgel determined by the use of TEM (transmission electron microscopy) tomography. The results show that the casein microgel displays a nonspecific attractive interaction for all macromolecules studied. When the macromolecular probes are spherical, this affinity is clearly size-dependent, with stronger attraction for the larger probes. The current data show that electrostatic effects cannot account for such an attraction. Rather, nonspecific hydration molecular forces appear to explain these results. These findings show how weak nonspecific forces affect the diffusion and partitioning of proteins and polysaccharides in a dense protein environment. These results could be useful to better understand the mechanisms of diffusion and partitioning in other media such as cells and tissues. Furthermore, there arises the possibility of using the casein micelle as a size-selective molecular device.

The Human Proteome Organisation's Proteomics Standards Initiative has developed the GelML (gel electrophoresis markup language) data exchange format for representing gel electrophoresis experiments performed in proteomics investigations. The format closely follows the reporting guidelines for gel electrophoresis, which are part of the Minimum Information About a Proteomics Experiment (MIAPE) set of modules. GelML supports the capture of metadata (such as experimental protocols) and data (such as gel images) resulting from gel electrophoresis so that laboratories can be compliant with the MIAPE Gel Electrophoresis guidelines, while allowing such data sets to be exchanged or downloaded from public repositories. The format is sufficiently flexible to capture data from a broad range of experimental processes, and complements other PSI formats for MS data and the results of protein and peptide identifications to capture entire gel-based proteome workflows. GelML has resulted from the open standardisation process of PSI consisting of both public consultation and anonymous review of the specifications.

One of the major safety issues in coal mining is heatings and the resultant spontaneous combustion in underground coal mines.CSIRO researchers have developed a number of polymer gels suitable for controlling heatings in coal mines.These gels were developed to meet strict selection criteria including easy preparation,no or low toxicity,controllable gelation time,adaptable to mine water chemistry,adjustable viscosity,relatively long gel life,thermally and chemically stable and low cost.The HPAM-Aluminum Citrate gel system was identified to be the most favourable gel system for fire suppression in underground coal mines.These gels can be applied to the areas undergoing coal heating or gas leakage at a controllable gelation time and impermeable gel barriers can be formed in the areas to block ingress of air.

Full Text Available Colloidal gels are possible intermediates in the generation of highly porous particle systems. In the production process the gels are fragmented after their formation. These gel fragments compact to particles whose application-technological properties are determined by their size and porosity. In the case of precipitated silica gels, this consolidation process depends on temperature and pH, among other parameters. It is shown that these dependencies can be characterized by oedometer measurements. Originally, the oedometer test (one-dimensional compression test stemmed from soil mechanics. It has proven to be an interesting novel examination method for gels. Quantitative data of the time-dependent shrinkage of gel samples can be obtained. The consolidation of the gels shows a characteristic dependence on the above parameters.

During lactation, mammary epithelial cells secrete huge amounts of milk from their apical side. The current view is that caseins are secreted by exocytosis, whereas milk fat globules are released by budding, enwrapped by the plasma membrane. Owing to the number and large size of milk fat globules, the membrane surface needed for their release might exceed that of the apical plasma membrane. A large-scale proteomics analysis of both cytoplasmic lipid droplets and secreted milk fat globule membranes was used to decipher the cellular origins of the milk fat globule membrane. Surprisingly, differential analysis of protein profiles of these two organelles strongly suggest that, in addition to the plasma membrane, the endoplasmic reticulum and the secretory vesicles contribute to the milk fat globule membrane. Analysis of membrane-associated and raft microdomain proteins reinforces this possibility and also points to a role for lipid rafts in milk product secretion. Our results provide evidence for a significant contribution of the endoplasmic reticulum to the milk fat globule membrane and a role for SNAREs in membrane dynamics during milk secretion. These novel aspects point to a more complex model for milk secretion than currently envisioned.

Kinesin-1 is a plus-end microtubule-based motor, and defects in kinesin-based transport are linked to diseases including neurodegeneration. Kinesin can auto-inhibit via a head-tail interaction, but is believed to be active otherwise. Here we report a tail-independent inactivation of kinesin, reversible by the disease-relevant signaling protein, casein kinase 2 (CK2). The majority of initially active kinesin (native or tail-less) loses its ability to interact with microtubules in vitro, and CK2 reverses this inactivation (~ 4-fold) without altering kinesin’s single motor properties. This activation pathway does not require motor phosphorylation, and is independent of head-tail auto-inhibition. In cultured mammalian cells, reducing CK2 expression, but not its kinase activity, decreases the force required to stall lipid droplet transport, consistent with a decreased number of active kinesin motors. Our results provide the first direct evidence of a protein kinase up-regulating kinesin-based transport, and suggest a novel pathway for regulating the activity of cargo-bound kinesin. PMID:22453827

Casein kinase G (CKG) with more than 2500-fold enrichraent was purified from Bufo bufo gargarizans ovaries. The catalytic activity of the enzyme was found to be associated with its 42 kD subunit, and its 26 kD subunit was found to be the major tsrget for the enzyme autophos phorylation. Each fuU-grown oocyte contained 1.9 units of CKG corresponding to an intracellular concentration of 93 nM. After injecting an amount of 0,38 units of the enzyme into the oocyte, approximately 50% of the progesterone-induoed maturation was inhibited. The inhibitory effect was enhanced in oocytes pretreated with spermine, which was consistent with the results that the enzyme was activated in vitro in the presence of spermine, The MPF-induced oocyte maturation was delayed and even prohibited in the kinase-microinjected oocytes. A 55 kD oocyte protein was identified as an suhstrate of CKG both in vivo and in vitro, and the enhancement of the 55 kD protein phosphory[ation was associated with kinase inhibition on maturation and on protein synthesis in kinase-microinjected oocytes. As the endogenous spermine level decreased in the course of progesteroneinduced oocyte maturation. 55 kD protein was dephosphorylated, Heparin, a specific inhibitor of CKG, potentiated the progesterone-induced oocyte maturation. Altogether the experimental reSults indicated Strongly that CKG may be the physiological target of spermine.

The Ikaros gene encodes a zinc finger,DNA-binding protein that regulates gene transcription and chromatin remodeling.Ikaros is a master regulator of hematopoiesis and an established tumor suppressor.Moderate alteration of Ikaros activity (e.g.haploinsufficiency) appears to be sufficient to promote malignant transformation in human hematopoietic cells.This raises questions about the mechanisms that normally regulate Ikaros function and the potential of these mechanisms to contribute to the development of leukemia.The focus of this review is the regulation of Ikaros function by phosphorylation/dephosphorylation.Site-specific phosphorylation of Ikaros by casein kinase 2 (CK2) controls Ikaros DNA-binding ability and subcellular localization.As a consequence,the ability of Ikaros to regulate cell cycle progression,chromatin remodeling,target gene expression,and thymocyte differentiation are controlled by CK2.In addition,hyperphosphorylation of Ikaros by CK2 leads to decreased Ikaros levels due to ubiquitinmediated degradation.Dephosphorylation of Ikaros by protein phosphatase 1 (PP1) acts in opposition to CK2 to increase Ikaros stability and restore Ikaros DNA binding ability and pericentromeric localization.Thus,the CK2 and PP1 pathways act in concert to regulate Ikaros activity in hematopoiesis and as a tumor suppressor.This highlights the importance of these signal transduction pathways as potential mediators of leukemogenesis via their role in regulating the activities of Ikaros.

Circadian clock coordinates behavior and physiology in mammals in response to light and feeding cycles. Disruption of normal clock function is associated with increased risk for cardiovascular and metabolic diseases, underscoring the emerging concept that temporal regulation of tissue metabolism is a fundamental aspect of energy homeostasis. We have previously demonstrated that transcriptional coactivator, peroxisome proliferator-activated receptor γ coactivator 1α (PGC-1α), coordinates circadian metabolic rhythms through simultaneous regulation of metabolic and clock gene expression. In this study, we found that PGC-1α physically interacts with, and is phosphorylated by, casein kinase 1δ (CK1δ), a core component of the circadian pacemaker. CK1δ represses the transcriptional function of PGC-1α in cultured hepatocytes, resulting in decreased gluconeogenic gene expression and glucose secretion. At the molecular level, CK1δ phosphorylation of PGC-1α within its arginine/serine-rich domain enhances its degradation through the proteasome system. Together, these results elucidate a novel mechanism through which circadian pacemaker transduces timing signals to the metabolic regulatory network that controls hepatic energy metabolism.

The aim of the present study was to evaluate the use of thermally-dried immobilized kefir on casein as a starter culture for protein-enriched dried whey cheese. For comparison reasons, dried whey cheese with thermally-dried free kefir culture and with no starter culture were also produced. The effect of the nature of the culture, the ripening temperature and the ripening process on quality characteristics of the whey cheese was studied. The association of microbial groups during cheese maturation suggested repression of spoilage and protection from pathogens due to the thermally-dried kefir, as counts of coliforms, enterobacteria and staphylococci were significantly reduced in cheeses produced using thermally-dried kefir starter cultures. The effect of the starter culture on production of volatile compounds responsible for cheese flavor was also studied using the SPME GC/MS technique. Thermally-dried immobilized kefir starter culture resulted in an improved profile of aroma-related compounds. The preliminary sensory evaluation ascertained the soft, fine taste and the overall improved quality of cheese produced with the thermally-dried immobilized kefir. The potential of protein-based thermally-dried starter cultures in dairy products is finally highlighted and assessed.

an altered sedimentation coefficient. The holoenzymes reconstituted with substituted mutants beta A 55,57, beta A55-57, and, to a lesser extent, beta A 59-61, beta A63,64, and beta A5,6 display a basal activity which is higher (up to 4-fold) than that of the wild type holoenzyme.(ABSTRACT TRUNCATED AT 250......Twenty-one mutants of the noncatalytic beta-subunit of human casein kinase-2 have been created, expressed in Escherichia coli, and purified to homogeneity. They are either modified at the autophosphorylation site (mutants beta delta 1-4 and beta A 5,6) or bear variable deletions in their C......-terminal part (mutants beta delta 209-215, beta delta 194-215, beta delta 181-215, beta delta 171-215, beta delta 150-215) or have undergone Ala substitutions for the acidic and basic residues which are concentrated in the sequences 55-70 and 171-180, respectively. All these mutants have been examined...

Complex sphingolipids are important components of eukaryotic cell membranes and, together with their biosynthetic precursors, including sphingoid long chain bases and ceramides, have important signaling functions crucial for cell growth and survival. Ceramides are produced at the endoplasmic reticulum (ER) membrane by a multicomponent enzyme complex termed ceramide synthase (CerS). In budding yeast, this complex is composed of two catalytic subunits, Lac1 and Lag1, as well as an essential regulatory subunit, Lip1. Proper formation of ceramides by CerS has been shown previously to require the Cka2 subunit of casein kinase 2 (CK2), a ubiquitous enzyme with multiple cellular functions, but the precise mechanism involved has remained unidentified. Here we present evidence that Lac1 and Lag1 are direct targets for CK2 and that phosphorylation at conserved positions within the C-terminal cytoplasmic domain of each protein is required for optimal CerS activity. Our data suggest that phosphorylation of Lac1 and Lag1 is important for proper localization and distribution of CerS within the ER membrane and that phosphorylation of these sites is functionally linked to the COP I-dependent C-terminal dilysine ER retrieval pathway. Together, our data identify CK2 as an important regulator of sphingolipid metabolism, and additionally, because both ceramides and CK2 have been implicated in the regulation of cancer, our findings may lead to an enhanced understanding of their relationship in health and disease. PMID:25429105

Complex sphingolipids are important components of eukaryotic cell membranes and, together with their biosynthetic precursors, including sphingoid long chain bases and ceramides, have important signaling functions crucial for cell growth and survival. Ceramides are produced at the endoplasmic reticulum (ER) membrane by a multicomponent enzyme complex termed ceramide synthase (CerS). In budding yeast, this complex is composed of two catalytic subunits, Lac1 and Lag1, as well as an essential regulatory subunit, Lip1. Proper formation of ceramides by CerS has been shown previously to require the Cka2 subunit of casein kinase 2 (CK2), a ubiquitous enzyme with multiple cellular functions, but the precise mechanism involved has remained unidentified. Here we present evidence that Lac1 and Lag1 are direct targets for CK2 and that phosphorylation at conserved positions within the C-terminal cytoplasmic domain of each protein is required for optimal CerS activity. Our data suggest that phosphorylation of Lac1 and Lag1 is important for proper localization and distribution of CerS within the ER membrane and that phosphorylation of these sites is functionally linked to the COP I-dependent C-terminal dilysine ER retrieval pathway. Together, our data identify CK2 as an important regulator of sphingolipid metabolism, and additionally, because both ceramides and CK2 have been implicated in the regulation of cancer, our findings may lead to an enhanced understanding of their relationship in health and disease.

Full Text Available This paper studies the properties of active polymer food packaging bilayer polyethylene/polycaprolactone (PE/PCL films. Such packaging material consists of primary PE layer coated with thin film of PCL coating modified with active component (zinc oxide or zinc oxide/casein complex with intention to extend the shelf life of food and to maintain the quality and health safety. The influence of additives as active components on barrier, mechanical, thermal and antimicrobial properties of such materials was studied. The results show that, in comparison to the neat PE and PE/PCL films, some of PE/PCL bilayer films with additives exhibit improved barrier properties i.e. decreased water vapour permeability. Higher thermal stability of modified PE/PCL material is obtained due to a modified mechanism of thermal degradation. The samples with the additive nanoparticles homogeneously dispersed in the polymer matrix showed good mechanical properties. Addition of higher ZnO content contributes to the enhanced antibacterial activity of a material.

Goat's αS1-casein (CSN1S1) polymorphism has a significant effect on milk protein and lipid composition, which affects the nutritional quality and technological properties of milk. Moreover, this polymorphism has a large impact on the morphology of mammary epithelial cells. To explore the metabolic pathways modulated in relation to this polymorphism, we compared the mammary gene expression profiles of two groups of lactating goats carrying either two reference or two defective alleles, using a bovine oligonucleotide microarray representing 8379 genes. We identified 41 differentially expressed genes between the two genotype groups. In particular, we showed a downregulation of two key lipogenic genes encoding fatty acid synthase and glycerol-3-phosphate acyltransferase in agreement with the low fat concentration associated with CSN1S1 deficiency. In addition, this study highlights changes in the expression level of several genes known to influence membrane fluidity, cell-cell interaction or chromatin organization. Our results open up new fields of investigation on structural modifications associated with CSN1S1 deficiency that could affect mammary gland function.

The aim of the current work was to analyze, in the Sarda breed goat, genetic polymorphism within the casein genes and to assess their influence on milk traits. Genetic variants at the CSN1S1, CSN2, CSN1S2 and CSN3 gene loci were investigated using PCR-based methods, cloning and sequencing. Strong alleles prevailed at the CSN1S1 gene locus and defective alleles also were revealed. Null alleles were evidenced at each calcium-sensitive gene locus. At the CSN3 gene locus, we observed a prevalence of the CSN3 A and B alleles; the occurrence of rare alleles such as CSN3 B'', C, C', D, E and M; and the CSN3 S allele (GenBank KF644565) described here for the first time in Capra hircus. Statistical analysis showed that all genes, except CSN3, significantly influenced milk traits. The CSN1S1 BB and AB genotypes were associated with the highest percentages of protein (4.41 and 4.40 respectively) and fat (5.26 and 5.34 respectively) (P milk protein content and yield. The polymorphism of the CSN2 gene affected milk protein percentage with the highest values recorded in the CSN2 AA goats (4.35, at P goats provided the highest fat (51.02 g/day) and protein (41.42 g/day) (P goat.

The concentration of casein (CN) is one of the most important parameters for measuring the quality of bovine milk. Traditional approach to CN concentration determination is Kjeldahl, which is an indirect method for determination of total nitrogen content. Here, we described a double-antibody based direct immunoassay for the detection of β-CN in bovine milk samples. Monoclonal antibody (McAb) was used as capture antibody and polyclonal antibody (PcAb) labelled with horseradish peroxidase (HRP) as detection antibody. With the direct immunoassay format, the linear range of the detection was 0.1-10.0 μg mL(-1). The detection limit was 0.04 μg mL(-1). In addition, the concentration of β-CN in real bovine milk samples has been detected by the developed immunoassay. There was a good correlation between the results obtained by the developed technique and Kjeldahl method from commercial samples. Compared to the traditional approach, the advantage of the assay is no need of time-consuming sample pretreatment.

Connexin (Cx) 45.6, an avian counterpart of rodent Cx50, is phosphorylated in vivo, but the sites and function of the phosphorylation have not been elucidated. Our peptide mapping experiments showed that the Ser(363) site in the carboxyl (COOH) terminus of Cx45.6 was phosphorylated and that this site is within casein kinase (CK) II consensus sequence, although showing some similarity to CKI sequence. The peptide containing Ser(363) could be phosphorylated in vitro by CKII, but not by CKI. Furthermore, CKII phosphorylated Cx45.6 in embryonic lens membrane and the fusion protein containing the COOH terminus of Cx45.6. Two-dimensional peptide mapping experiments showed that one of the Cx45.6 peptides phosphorylated in vivo migrated to the same spot as one of those phosphorylated by CKII in vitro. Furthermore, CKII activity could be detected in lens lysates. To assess the function of this phosphorylation event, exogenous wild type and mutant Cx45.6 (Ser(363) --> Ala) were expressed in lens primary cultures by retroviral infection. The mutant Cx45.6 was shown to be more stable having a longer half-life compared with wild type Cx45.6. Together, the evidence suggests that CKII is likely a kinase responsible for the Ser(363) phosphorylation, leading to the destablization and degradation of Cx45.6. The connexin degradation induced by phosphorylation has a broad functional significance in the regulation of gap junctions in vivo.

This work reports on the development and characterization of novel meltable polymers and composites based on casein and soybean proteins. The effects of inert (Al(2)O(3)) and bioactive (tricalcium phosphate) ceramic reinforcements over the mechanical performance, water absorption, and bioactivity behavior of the injection-molded thermoplastics were examined. It was possible to obtain materials and composites with a range of mechanical properties, which might allow for their application in the biomedical field. The incorporation of tricalcium phosphate into the soybean thermoplastic decreased its mechanical properties but lead to the nucleation of a bioactive calcium-phosphate film on their surface when immersed in a simulated body fluid solution. When compounded with 1% of a zirconate coupling agent, the nucleation and growth of the bioactive films on the surface of the referred to composites was accelerated. The materials degradation was studied for ageing periods up to 60 days in an isotonic saline solution. Both water uptake and weight loss were monitored as a function of the immersion time. After 1 month of immersion, the materials showed signal of chemical degradation, presenting weight losses up to 30%. However, further improvement on the mechanical performance and the enhancement of the hydrolytic stability of those materials will be highly necessary for applications in the biomedical field.

Full Text Available Plasmopara viticola, the causal agent of grapevine downy mildew, is one of the most devastating grape pathogen in Europe and North America. Although phytochemicals are used to control pathogen infections, the appearance of resistant strains and the concern for possible adverse effects on environment and human health are increasing the search for alternative strategies. In the present investigation, we successfully tested two protein hydrolysates from soybean (soy and casein (cas to trigger grapevine resistance against P. viticola. On Vitis vinifera cv. Marselan plants, the application of soy and cas reduced the infected leaf surface by 76 and 63%, as compared to the control, respectively. Since both hydrolysates might trigger the plant immunity, we investigated their ability to elicit grapevine defence responses. On grapevine cell suspensions, a different free cytosolic calcium signature was recorded for each hydrolysate, whereas a similar transient phosphorylation of two MAP kinases of 45 and 49 kDa was observed. These signalling events were followed by transcriptome reprogramming, including the up-regulation of defence genes encoding pathogenesis-related (PR proteins and the stilbene synthase enzyme responsible for the biosynthesis of resveratrol, the main grapevine phytoalexin. Liquid chromatography analyses confirmed the production of resveratrol and its dimer metabolites, δ- and ε-viniferins. Overall, soy effects were more pronounced as compared to the cas one. Both hydrolysates proved to act as elicitors to enhance grapevine immunity against pathogen attack.

Full Text Available The Earth's rotation has driven the evolution of cellular circadian clocks to facilitate anticipation of the solar cycle. Some evidence for timekeeping mechanism conserved from early unicellular life through to modern organisms was recently identified, but the components of this oscillator are currently unknown. Although very few clock components appear to be shared across higher species, Casein Kinase 1 (CK1 is known to affect timekeeping across metazoans and fungi, but has not previously been implicated in the circadian clock in the plant kingdom. We now show that modulation of CK1 function lengthens circadian rhythms in Ostreococcustauri, a unicellular marine algal species at the base of the green lineage, separated from humans by ~1.5 billion years of evolution. CK1 contributes to timekeeping in a phase-dependent manner, indicating clock-mediated gating of CK1 activity. Label-free proteomic analyses upon overexpression as well as inhibition revealed CK1-responsive phosphorylation events on a set of target proteins, including highly conserved potentially clock-relevant cellular regulator proteins. These results have major implications for our understanding of cellular timekeeping and can inform future studies in any circadian organism.

Full Text Available Throughout the day, clock proteins synchronize changes in animal physiology (e.g., wakefulness and appetite with external cues (e.g., daylight and food. In vertebrates, both casein kinase 1 delta and epsilon (CK1δ and CK1ε regulate these circadian changes by phosphorylating other core clock proteins. In addition, CK1 can regulate circadian-dependent transcription in a non-catalytic manner, however, the mechanism is unknown. Furthermore, the extent of functional redundancy between these closely related kinases is debated. To further advance knowledge about CK1δ and CK1ε mechanisms of action in the biological clock, we first carried out proteomic analysis of both kinases in human cells. Next, we tested interesting candidates in a cell-based circadian readout which resulted in the discovery of PROHIBITIN 2 (PHB2 as a modulator of period length. Decreasing the expression of PHB2 increases circadian-driven transcription, thus revealing PHB2 acts as an inhibitor in the molecular clock. While stable binding of PHB2 to either kinase was not detected, knocking down CK1ε expression increases PHB2 protein levels and, unexpectedly, knocking down CK1δ decreases PHB2 transcript levels. Thus, isolating CK1 protein complexes led to the identification of PHB2 as an inhibitor of circadian transcription. Furthermore, we show that CK1δ and CK1ε differentially regulate the expression of PHB2.

Traditional Kjeldahl method, used for quality evaluation of bovine milk, has intrinsic defects of time-consuming sample preparation and two analyses to determine the difference between non-protein nitrogen content and total protein nitrogen content. Herein, based upon antibody functionalized gold nanoparticles (AuNPs), we described a colorimetric method for β-casein (β-CN) detection in bovine milk samples. The linear dynamic range and the LOD were 0.08-250 μg mL(-1), and 0.03 μg mL(-1) respectively. In addition, the real content of β-CN in bovine milk was measured by using the developed assay. The results are closely correlated with those from Kjeldahl method. The advantages of β-CN triggered AuNP aggregation-based colorimetric assay are simple signal generation, the high sensitivity and specificity as well as no need of complicated sample preparation, which make it for on-site detection of β-CN in bovine milk samples.

A novel particulate delivery matrix based on ionically crosslinked casein (CAS) nanoparticles was developed for controlled release of the poorly soluble anticancer drug flutamide (FLT). Nanoparticles were fabricated via oil-in-water emulsification then stabilized by ionic crosslinking of the positively charged CAS molecules below their isoelectric point, with the polyanionic crosslinker sodium tripolyphosphate. With the optimal preparation conditions, the drug loading and incorporation efficiency achieved were 8.73% and 64.55%, respectively. The nanoparticles exhibited a spherical shape with a size below 100 nm and a positive zeta potential (+7.54 to +17.3 mV). FLT was molecularly dispersed inside the nanoparticle protein matrix, as revealed by thermal analysis. The biodegradability of CAS nanoparticles in trypsin solution could be easily modulated by varying the sodium tripolyphosphate crosslinking density. A sustained release of FLT from CAS nanoparticles for up to 4 days was observed, depending on the crosslinking density. After intravenous administration of FLT-CAS nanoparticles into rats, CAS nanoparticles exhibited a longer circulation time and a markedly delayed blood clearance of FLT, with the half-life of FLT extended from 0.88 hours to 14.64 hours, compared with drug cosolvent. The results offer a promising method for tailoring biodegradable, drug-loaded CAS nanoparticles as controlled, long-circulating drug delivery systems of hydrophobic anticancer drugs in aqueous vehicles.

Throughout the day, clock proteins synchronize changes in animal physiology (e.g., wakefulness and appetite) with external cues (e.g., daylight and food). In vertebrates, both casein kinase 1 delta and epsilon (CK1δ and CK1ε) regulate these circadian changes by phosphorylating other core clock proteins. In addition, CK1 can regulate circadian-dependent transcription in a non-catalytic manner, however, the mechanism is unknown. Furthermore, the extent of functional redundancy between these closely related kinases is debated. To further advance knowledge about CK1δ and CK1ε mechanisms of action in the biological clock, we first carried out proteomic analysis of both kinases in human cells. Next, we tested interesting candidates in a cell-based circadian readout which resulted in the discovery of PROHIBITIN 2 (PHB2) as a modulator of period length. Decreasing the expression of PHB2 increases circadian-driven transcription, thus revealing PHB2 acts as an inhibitor in the molecular clock. While stable binding of PHB2 to either kinase was not detected, knocking down CK1ε expression increases PHB2 protein levels and, unexpectedly, knocking down CK1δ decreases PHB2 transcript levels. Thus, isolating CK1 protein complexes led to the identification of PHB2 as an inhibitor of circadian transcription. Furthermore, we show that CK1δ and CK1ε differentially regulate the expression of PHB2.

Experiments were conducted with young chicks to quantify the absorption efficiency of manganese (Mn) using tibia Mn uptake as the response criterion. Chicks in experiment 1 were fed a corn-soybean meal diet and either injected intraperitoneally or crop intubated twice daily with three levels of inorganic Mn provided as MnSO4 X H2O. After 14 d, chicks were killed, and intact tibias were removed to facilitate calculation of bone Mn concentration. Excellent straight-line fits (bone Mn concentration as a function of Mn administered) allowed estimation of gut absorption efficiency of Mn by slope-ratio methodology. Absorption efficiency of Mn was calculated to be 1.71% for chicks fed the corn-soy diet. Identical methodology was employed in experiment 2 wherein chicks were fed a phytate- and fiber-free casein-dextrose diet. An absorption efficiency of 2.40% was estimated for chicks fed this diet. Hence, absorption efficiency of inorganic Mn was 40% greater in chicks fed the purified diet than in those fed the conventional corn-soy diet.

Meat and bone meal (MBM) is a high protein agricultural commodity that currently has few applications other than as an animal feed. Unmodified MBM has poor functional properties, due to its low solubility. Our results from pilot plant trials demonstrate that MBM can be extrusion-processed along with sodium caseinate to produce a useful plastic material. We developed this material for use as a dog chew toy. For this application, elastic modulus (stiffness) is a key characteristic. Our results detail the relationship between ambient relative humidity and equilibrium moisture content (MC) in the material. The influence of MC on the glass transition temperature and elastic modulus reflects the plasticization of this material by water. On the basis of a comparison to a commercially available dog chew, the range of stiffness achievable with our material, 0.25-2.50 GPa, encompasses the values appropriate for a dog chew. Our results show that a particular desired stiffness can be maintained by applying an edible moisture barrier to the surface of the material.

of wine as clarification processing aids pursuant to Article 6, paragraph 11 of Directive 2000/13/EC – for permanent exemption from labelling. In earlier assessments, the Panel concluded that wines fined with casein/caseinate/milk products may trigger adverse reactions in susceptible individuals under...... refers to new analytical methods developed for the detection of milk allergens in the fining agent and the detection of casein in wine. There were no changes in the wine manufacturing process and no new clinical studies were provided. Taking into account the information provided on the characterisation...

enzyme) showed identical activity and the same molecular weight. On sodium dodecyl sulphate/polyacrylamide-gel electrophoresis a single band of about 70 000 mol.wt. was observed. Sucrose-gradient analysis, however, showed that the enzyme activity sedimented with a s20,w of approx. 7.5S. This observation...

Caseins are widely used for species identification of dairy products. Isoelectric focusing (IEF) of para-κ-casein peptide is used as the official German method for the differentiation between caprine (isoform A) and ovine (isoform B) dairy products, based on their different isoelectric points. The discrimination between Greek goat and ewe dairy products using IEF has, however, been shown to be problematic because of the existence of the ewe isoform in milk from Greek indigenous dairy goats. This could be due to nucleotide polymorphisms within the goat κ-casein gene of Greek indigenous breeds, which alter the isoelectric point of the para-κ-casein peptide and lead to false positive results. Previous DNA analysis of the goat κ-casein gene has shown high levels of polymorphism; however, no such information is available for Greek indigenous dairy goats. Therefore, 87 indigenous dairy goats were sequenced at exon IV of κ-casein gene. In total, 9 polymorphic sites were detected. Three nonsynonymous point mutations were identified, which change the isoelectric point of the goat para-κ-casein peptide so that it appears identical to that of the ewe peptide. Ten composite genotypes were reconstructed and 6 of them included the problematic point mutations. For the verification of genetic results, IEF was carried out. Both goat and ewe patterns appeared in the problematic genotypes. The frequency of these genotypes could be characterized as moderate (0.23) to high (0.60) within Greek indigenous breeds. However, this is not an issue restricted to Greece, as such genotypes have been detected in various non-Greek goat breeds. In conclusion, IEF based on the official German method is certainly inappropriate for ovine and caprine discrimination concerning Greek dairy goat products, and consequently a new method should be established.

The present invention relates to a single phase metal-silica sol-gel glass formed by the co-condensation of a transition metal with silicon atoms where the metal atoms are uniformly distributed within the sol-gel glass as individual metal centers. Any transition metal may be used in the sol-gel glasses. The present invention also relates to sensor materials where the sensor material is formed using the single phase metal-silica sol-gel glasses. The sensor materials may be in the form of a thin film or may be attached to an optical fiber. The present invention also relates to a method of sensing chemicals using the chemical sensors by monitoring the chromatic change of the metal-silica sol-gel glass when the chemical binds to the sensor. The present invention also relates to oxidation catalysts where a metal-silica sol-gel glass catalyzes the reaction. The present invention also relates to a method of performing oxidation reactions using the metal-silica sol-gel glasses. The present invention also relates to organopolymer metal-silica sol-gel composites where the pores of the metal-silica sol-gel glasses are filled with an organic polymer polymerized by the sol-gel glass.

All solid state electrochromic devices have potential applications in architectural and automotive fields to regulate the transmission and reflection of radiant energy. We present the optical and electrochemical characteristics of two solid state windows having the configuration glass/ITO/TiO2-CeO2/TiO2/TiO2-CeO2/ITO/glass and glass/ITO/WOa/TiO2/TiO2-CeO2/ITO/glass where the three internal layers have been prepared by sol gel methods. The preparation of the individual sols and some physical p...

Motility initiation in crawling cells requires a symmetry breaking mechanism which transforms a symmetric state into a polarized state. Experiments on keratocytes suggest that polarization is triggered by increased contractility of motor proteins. In this paper we argue that contraction can be responsible not only for the symmetry breaking transition but also for the incipient translocation of the segment of an active gel mimicking the crawling cell. Our model suggests that when the contractility increases sufficiently far beyond the motility initiation threshold, the cell can stop and re-symmetrizes. The proposed theory reproduces the motility initiation pattern in fish keratocytes and the behavior of keratocytes prior to cell division.

Heat (85 degrees C for 20 min) and pressure (600 MPa for 15 min) treatments were applied to skim milk fortified by addition of whey protein concentrate. Both treatments caused > 90 % denaturation of beta-lactoglobulin. During heat treatment this denaturation took place in the presence of intact casein micelles; during pressure treatment it occurred while the micelles were in a highly dissociated state. As a result micelle structure and the distribution of beta-lactoglobulin were different in the two milks. Electron microscopy and immunolabelling techniques were used to examine the milks after processing and during their transition to yogurt gels. The disruption of micelles by high pressure caused a significant change in the appearance of the milk which was quantified by measurement of the colour values L*, a* and b*. Heat treatment also affected these characteristics. Casein micelles are dynamic structures, influenced by changes to their environment. This was clearly demonstrated by the transition from the clusters of small irregularly shaped micelle fragments present in cold pressure-treated milk to round, separate and compact micelles formed on warming the milk to 43 degrees C. The effect of this transition was observed as significant changes in the colour indicators. During yogurt gel formation, further changes in micelle structure, occurring in both pressure and heat-treated samples, resulted in a convergence of colour values. However, the microstructure of the gels and their rheological properties were very different. Pressure-treated milk yogurt had a much higher storage modulus but yielded more readily to large deformation than the heated milk yogurt. These changes in micelle structure during processing and yogurt preparation are discussed in terms of a recently published micelle model.

We report an enabling method for mapping the protein sequence with high sequence coverage. This method combines the high separation power of gel electrophoresis for protein separation with the high sequence coverage capability of microwave-assisted acid hydrolysis (MAAH) mass spectrometry (MS). In-gel MAAH using 25% trifluoroacetic acid was developed and optimized for degrading the gel-separated protein into small peptides suitable for tandem MS sequencing. For bovine serum albumin (BSA) (∼67 kDa), with 4 μg of protein loading onto a gel for separation, followed by excising the protein gel band for in-gel MAAH and then injecting ∼2 μg of the resultant peptides into a liquid chromatography quadrupole time-of-flight mass spectrometer for analysis, 689 ± 54 (n = 3) unique peptides were identified with a protein sequence coverage of 99 ± 1%. Both the number of peptides detected and sequence coverage decreased as the sample amount decreased, mainly due to background interference: 316 ± 59 peptides and 94 ± 3% coverage for 2 μg loading, 136 ± 19 and 76 ± 5% for 1 μg loading, and 30 ± 2 and 32 ± 2% for 0.5 μg loading. To demonstrate the general applicability of the method, 10 gel bands from gel electrophoresis of an albumin-depleted human plasma sample were excised for in-gel MAAH LC-MS analysis. In total, 19 relatively high abundance proteins with molecular weights ranging from ∼8 to ∼160 kD could be mapped with coverage of 100% for six proteins (MW 8759 to 68 425 Da), 96-98% for five proteins (MW 11 458 to 36 431 Da), 92% for three proteins (MW 15 971 to 36 431 Da), 80-87% for four proteins (MW 42 287 to 162 134 Da), and 56% for one protein (MW 51 358 Da). Finally, to demonstrate the applicability of the method for more detailed analysis of complex protein mixtures, two-dimensional (2D) gel electrophoresis was combined with in-gel MAAH, affinity purification, and LC-MS/MS to characterize six bovine alpha-S1-casein phosphoprotein

We develop a model of amoeboid cell motility based on active gel theory. Modeling the motile apparatus of a eukaryotic cell as a confined layer of finite length of poroelastic active gel permeated by a solvent, we first show that, due to active stress and gel turnover, an initially static and homogeneous layer can undergo a contractile-type instability to a polarized moving state in which the rear is enriched in gel polymer. This agrees qualitatively with motile cells containing an actomyosin-rich uropod at their rear. We find that the gel layer settles into a steadily moving, inhomogeneous state at long times, sustained by a balance between contractility and filament turnover. In addition, our model predicts an optimal value of the gel-susbstrate adhesion leading to maximum layer speed, in agreement with cell motility assays. The model may be relevant to motility of cells translocating in complex, confining environments that can be mimicked experimentally by cell migration through microchannels.

Isotropic active gels are the network which is consist of cross-linked building blocks and the structure of which changes randomly and isotropically with time. Dogic et. al. show that pairs of anti-parallel microtubules form extensile bundles, which merge, extend, and buckle. In an unconfined system, the dynamics of these bundles causes spontaneous turbulent-like flow driven by motion of microscopic molecular motors. We found that confining these active gels in a millimeter sized toroids causes a transition into a new dynamical state characterized by circulation currents persisting for hours until ATP is depleted. We show how toroid dimensions impact the properties of self-organized circular currents, how directions of circulation can be designed by engineering ratchet-shaped boundaries, and how circulations of connected toroids can be either synchronized or antisynchronized. Furthermore, we demonstrate that the flow rate in the circulation is independent of curvature and length of flow path. The flow rate persists for centimeters without decay, disregarding conventional pipe flow resistance. Such findings pave the path to self-pumping pipe transport and performing physical work with biological system.

Phase density differences in sols, foams, or emulsions often lead to sedimentation or creaming, causing problems for materials where spatial uniformity over extended periods of time is essential. The problem may be addressed through the use of rheology modifiers in the continuous phase. Weak polymer gels have found use for this purpose in the food industry where they appear to be capable of trapping dispersoid particles in a three-dimensional matrix while displaying water-like viscosities at low shear. Attempts to predict sedimentation stability in terms of particle properties (size, shape, density difference) and gel yield stress have led to qualitative success for suspensions of large particles. The effect of particle size, however, in particular the case in which colloidal dimensions are approached, has not been investigated. The present work seeks to determine useful stability criteria for colloidal dispersions in terms of readily accessible viscoelastic descriptors. Results are reported for systems consisting of 12 microm poly(methyl methacrylate) (PMMA) spheres dispersed in aqueous gellan gum. Monovalent salt concentration is varied to control rheological properties, and sedimentation/centrifugation experiments are performed to determine dispersion stability. Necessary conditions for stability consist of a minimum yield stress together with a value of tan delta less than unity.

The objective of this work was to examine whether variation in the amino acid supply to cows could be a reason for the reduced casein content and poorer renneting properties of milk that often occur in late summer, or whether these effects are related to proteolysis in the raw milk. In a 2 x 2 x 2 factorial design, we investigated the effects of sward (clover v. rye-grass) and supplementary feed with a high or low level of rumen-soluble N or of rumen undegradable protein on milk protein composition during the grazing season. A total of 32 Danish Holstein cows were included in the experiment. Milk protein and casein contents and the ratios casein N:total N and casein:true protein were at a minimum in late summer, whereas the contents of urea, non-protein N and whey protein were higher during this period. These seasonal effects were unrelated to either the type of supplementary feed or the type of sward; neither were they clearly related to proteolysis, although casein: true protein was related to the proteose peptone content. The results indicated that whey proteins other than alpha-lactalbumin or beta-lactoglobulin accounted for the higher proportion or concentration of whey protein in late summer. Based on a principal component analysis including variables such as citric acid, lactose and non-protein N, we suggest that the cows' energy supply during this period may be a critical factor in determining the milk protein composition, although our results were not conclusive. There was an interaction between the supplement of rumen undegradable protein and type of sward. When clover was grazed, a high supplement increased the concentrations of protein and casein in milk and the kappa-casein: total casein ratio. When rye-grass was grazed, the opposite response was found, and overall milk protein yield was not affected. The very low N content of clover in early summer reduced milk protein and casein protein during this period.

The purpose of this study was to compare characteristics of radiochromic gel - Turnbull Blue gel (TB gel) with polymer gel - polyacrylamide gel and tetrakis hydroxymethyl phosphonium chloride (PAGAT) using optical tomography. Both types of gels were examined in terms of dose sensitivity, dose response linearity and background value of spectrophotometric absorbance. The calibration curve was obtained for 60Co irradiation performed on Gammacell 220 at predefined gamma dose levels between 0 and 140 Gy for TBG and 0-15 Gy for PAGAT. To measure relative dose distributions from stereotactic irradiation, dosimeters were irradiated on Leksell Gamma Knife Perfexion. The cylindrical glass housings filled with gel were attached to the stereotactic frame. They were exposed with single shot and 16 mm collimator by 65 Gy to a 50% prescription isodose for TB gel and 4 Gy to a 50% prescription isodose for PAGAT. Evaluations of dosimeters were performed on an UV-vis Spectrophotometer Helios β and an optical cone beam homemade tomography scanner with a 16-bit astronomy CCD camera with a set of color filters. The advantages and potential disadvantages for both types of gel dosimeters were summarized. Dose distribution in central slice and measured profiles of 16 mm shot shows excellent correspondence with treatment planning system Leksell GammaPlan® for both PAGAT and Turnbull Blue gels. Gel dosimeters are suitable for steep dose gradient verification. An optical tomography evaluation method is successful. Dose response characteristics of TB gel and PAGAT gel are presented.

High-dose chemotherapy and surgical treatment have improved the prognosis of osteosarcoma. However, more than 20% of patients with osteosarcoma still have a poor prognosis. We investigated the expression and function of casein kinase 2 (CK2) in osteosarcoma growth. We then examined the effects of CX-4945, a CK2 inhibitor, on osteosarcoma growth in vitro and in vivo to apply our findings to the clinical setting. We examined the expression of CK2α and CK2β by western blot analysis, and performed WST-1 assays using CK2α and CK2β siRNA or CX-4945. Flow cytometry and western blot analyses were performed to evaluate apoptotic cell death. Xenograft models were used to examine the effect of CX-4945 in vivo. Western blot analysis revealed upregulation of CK2α and CK2β in human osteosarcoma cell lines compared with human osteoblast cells or mesenchymal stem cells. WST assay showed that knockdown of CK2α or CK2β by siRNA inhibited the proliferation of human osteosarcoma cells. Treatment with 3 µM of CX-4945 inhibited osteosarcoma cell proliferation; however, the same concentration of CX-4945 did not affect the proliferation of human mesenchymal stem cells. Additionally, treatment with CX-4945 inhibited the proliferation of human osteosarcoma cells in a dose-dependent manner. Western blot and flow cytometry analyses showed that treatment with CX-4945 promoted apoptotic death of osteosarcoma cells. The xenograft model showed that treatment with CX-4945 significantly prevented osteosarcoma growth in vivo compared with control vehicle treatment. Our findings indicate that CK2 may be an attractive therapeutic target for treating osteosarcoma.

Trichoderma reesei is the most important industrial cellulase-producing filamentous fungus. Although its molecular physiology has been investigated, the signal transduction pathways are not fully understood. In particular, the role of casein kinase II (CKII) is not yet clear. In this work, we carried out functional investigations on a catalytic subunit of CKII, CKIIα2. Comparison of the phenotypic features of T. reesei parent and Δck2α2 strains showed significant changes following ck2α2 disruption. T. reesei Δck2α2 form significantly smaller mycelial pellets in glucose-containing liquid minimum media, have shorter and fewer branch hyphae, produce smaller amounts of chitinases, produce more spores, show more robust growth on glucose-containing agar plates, and consume glucose at a significantly higher rate. Suggestions can be made that CKIIα2 governs chitinase expression, and the disruption of ck2α2 results in lower levels of chitinase production, leading to a weaker cell wall disruption capability, further resulting in weaker hyphal branching, which eventually leads to smaller mycelial pellets in liquid media. Further conclusions can be made that CKIIα2 is involved in repression of sporulation and glucose metabolism, which is consistent with the proposal that CKIIα2 represses global metabolism. These observations make the deletion of ck2α2 a potentially beneficial genetic disruption for T. reesei during industrial applications, as smaller mycelial pellets, more spores and more robust glucose metabolism are all desired traits for industrial fermentation. This work reports novel unique functions of a CKII catalytic subunit and is also the first genetic and physiological investigation on CKII in T. reesei.

Background The gluten free, casein free (GFCF) diet is heralded by strong anecdotal parental reports to greatly improve and even “cure” symptoms of Autism Spectrum Disorders (ASD). Yet to date, little conclusive empirical evidence exists supporting its use. Objective The purpose of this paper is to provide an overview of the state of the recent evidence regarding use of GFCF diet for treatment of individuals with ASD. Methods Five database providers (PubMed, Web of Knowledge, EBSCO, ProQuest, and WorldCat) were used to search 19 databases yielding a total of 491 articles that were published through February 2015. Peer reviewed articles published between 2005 and February 2015 were included for review if study participants were identified as having ASD and investigated the effects of the GFCF diet on ASD behaviors or the relationship between the diet and these behaviors. Results Evaluation of search results yielded 11 reviews, 7 group experimental studies including 5 randomized controlled trials, 5 case reports, and 4 group observational studies published during the last 10 years. These studies represent a marked increase in number of reported studies as well as increased scientific rigor in investigation of GFCF diets in ASD. Conclusions While strong empirical support for the GFCF diet in ASD is currently lacking, studies point to the need for identifying subsets of individuals (e.g., those with documented gastrointestinal abnormalities) who may be the best responders to the GFCF diet. Identifying these subsets is critically needed to enhance rigor in this research area. Until rigorous research supporting use of GFCF diet is reported, clinicians should continue use caution and consider several factors when advising regarding implementation of the GFCF diet for individuals with ASD. PMID:28111520

This study was carried out to evaluate the difference between bonding to demineralized enamel and remineralized enamel using casein phosphopeptide-amorphous calcium phosphate with fluoride (CPP-ACFP) or without fluoride (CPP-ACP) compared to normal enamel. Another aim was to test if the newly introduced Single Bond Universal adhesive system would show better bonding to any enamel condition in comparison to the other tested adhesive systems. The lingual enamel surfaces of 40 non carious human third molars were divided into four main groups according to the enamel condition (ground normal enamel [negative control]; demineralized enamel [positive control]; and remineralized enamel with CPP-ACP or with CPP-ACFP, respectively). Within each main group, the lingual enamel surface of each tooth was sectioned into three slabs, resulting in 30 slabs that were distributed into three subgroups according to the adhesive system utilized (Clearfil S(3) Bond Plus, Single Bond Universal, or G-aenial Bond). Two resin composite microcylinder buildups were made on each enamel slab using Filtek Z350 XT. The μSBS was evaluated at a crosshead speed of 0.5 mm/min. Modes of failure were detected using an environmental scanning electron microscope at 300× magnification. The two-way analysis of variance with repeated measures revealed a significant effect for the enamel condition. However, there was no significant effect for the type of adhesive system. The interaction between the enamel condition and the type of adhesive system was also not significant. Modes of failure were mainly adhesive except for the demineralized enamel. It showed a mixed type of failure, in which cohesive failure in enamel was recorded. All single-step self-etch adhesives revealed comparable μSBS values to ground enamel and enamel remineralized with CPP-ACP or CPP-ACFP. Bonding to demineralized enamel was ineffective. With any enamel condition, no tested single-step self-etch adhesive was superior in its bonding.

Whey protein (WP) is characterized as a "fast" protein and caseinate (CA) as a "slow" protein according to their digestion and absorption rates. We hypothesized that co-ingestion of milk proteins (WP and CA) may be effective for prolonging the muscle protein synthesis response compared to either protein alone. We therefore compared the effect of ingesting milk protein (MP) to either WP or CA alone on muscle protein synthesis after exercise in rats. We also compared the effects of these milk-derived proteins to a control, soy protein (SP). Male Sprague-Dawley rats swam for two hours. Immediately after exercise, one of the following four solutions was administered: WP, CA, MP, or SP. Individual rats were euthanized at designated postprandial time points and triceps muscle samples collected for measurement of the protein fractional synthesis rate (FSR). FSR tended to increase in all groups post-ingestion, although the initial peaks of FSR occurred at different times (WP, peak time = 60 min, FSR = 7.76%/day; MP, peak time = 90 min, FSR = 8.34%/day; CA, peak time = 120 min, FSR = 7.85%/day). Milk-derived proteins caused significantly greater increases (p < 0.05) in FSR compared with SP at different times (WP, 60 min; MP, 90 and 120 min; CA, 120 min). Although statistical analysis could not be performed, the calculated the area under the curve (AUC) values for FSR following this trend were: MP, 534.61; CA, 498.22; WP, 473.46; and SP, 406.18. We conclude that ingestion of MP, CA or WP causes the initial peak time in muscle protein synthesis to occur at different times (WP, fast; MP, intermediate; CA, slow) and the dairy proteins have a superior effect on muscle protein synthesis after exercise compared with SP.

Calcium acrylate is a polymerized grout, and can polymerize in an aqueous solution. The polymerizationreaction utilizes ammonium persulfate as a catalyst and sodium thiosulfate as the activator. Based on the theory of reactionkinetics, this study on the relation between gel time and concentration of activator and catalyst showed that gel time ofcalcium acrylate is inversely proportional to activator and catalyst concentration. A formula of gel time is proposed, and anexample is provided to verify the proposed formula.

Calcium acrylate is a polymerized grout, and can polymerize in an aqueous solution. The polymerization reaction utilizes ammonium persulfate as a catalyst and sodium thiosulfate as the activator. Based on the theory of reaction kinetics, this study on the relation between gel time and concentration of activator and catalyst showed that gel time of calcium acrylate is inversely proportional to activator and catalyst concentration. A formula of gel time is proposed, and an example is provided to verify the proposed formula.

New improved aqueous gels, and methods of using same in contacting subterranean formations, are provided. The gels are prepared by gelling an aqueous brine having incorporated therein a water-soluble cellulose ether such as a carboxymethylcellulose (CMC), and are rendered more stable to decomposition by incorporating a sulfoalkylated tannin stabilizing agent, such as a sulfomethylated quebracho (SMQ), in the gel during the preparation thereof.

The objectives of this study were to determine the effects of a single gene and composite genotype of the casein gene family, including the beta-lactoglobulin gene (beta-LGB), acyl-CoA: diacylglycerol acyltransferase 1 gene (DGAT1), growth hormone gene (GH), and luteinizing hormone receptor gene (LHR) on milk yield, milk composition, the percentage of fat, protein, solids-not-fat, and total solid in crossbred Holsteins. A total of 231 crossbred Holstein cows were examined for the study. The genotype of the beta-casein gene was analyzed by allele-specific polymerase chain reaction, while the alpha-S1, alpha-S2, kappa-casein, DGAT1, beta-LGB, and GH genes were analyzed using a polymerase chain reaction-restriction fragment length polymorphism assay. The association between genes and milk yield and milk composition was analyzed. Three pairs of genes, for which significant associations were detected, were beta + kappa-casein, DGAT1 + beta-casein, and GH + beta-LGB. In the single-gene model, most loci are significantly associated with traits. A significant association between the composite genotype and the traits was detected in all composite genotypes. GH + beta-LGB appears to be the most suitable variants for improving milk production and percentage of milk protein. Overall, the effects of the composite genotype and single gene were different. A physical or functional relationship between genes is necessary for investigating gene markers.

Full Text Available Oxidation was related to the pathogenesis of human diseases. Adequate intake of antioxidant activity of food can reduce the levels of free radicals, prevent lipid peroxidation, and help the body against diseases. In the paper, casein from goat milk was hydrolyzed by five commercial proteases, namely, Alcalase, flavourzyme, papain, proteinase K and trypsin. The antioxidant activities of casein hydrolysates were assessed by evaluating hydrolysis degree, DPPH radical-scavenging activity, metal-chelating activity and superoxide radical scavenging activity. The results showed as follows: the DH of proteinase K, Alcalase, and trypsin were higher significantly than those of papain and flavourzyme. The Fe2+-chelation activity and superoxide radical scavenging activity of casein hydrolysates from goat milk by Alcalase was higher than the others, the DPPH scavenging activities of casein hydrolysates by Alcalase and papain was higher than the others and the DPPH scavenging activities by Alcalase and papain had no significant diffierence (p<0.05, so the optimal proteinase for hydrolysis casein from goat milk to produce antioxidant peptide was Alcalase.

A number of studies report that a diet high in protein influences cognitive performance, but the results are inconsistent. Studies demonstrated that protein from different food sources has differential effects on cognition. It is increasingly recognized that the integrity of cerebrovascular blood-brain barrier (BBB) is pivotal for central nervous system function. However, to date, no studies have reported the effects of high-protein diets on BBB integrity. Therefore, in this study, the effects of diets enriched in casein or soy protein on BBB permeability were investigated. Immunomicroscopy analyses of cerebral parenchymal immunoglobulin G extravasation indicated significant BBB disruption in the cortex of young adult mice maintained on high-casein diet for 12 weeks, while no signs of BBB dysfunction were observed in mice fed with control or high-soy protein diet. Moreover, cortical expression of glial fibrillary acidic protein (GFAP) was significantly greater in mice fed the high-casein diet compared to control mice, indicating heightened astrocyte activation, whereas mice maintained on a soy-enriched diet showed no increase of GFAP abundance. Plasma concentrations of homocysteine were markedly greater in mice maintained on a high-casein diet in comparison to control mice. Collectively, these findings suggest that a diet enriched in casein but not soy protein may induce astrocyte activation through exaggerated BBB permeability by increased plasma homocysteine. The outcomes indicate the differential effects of protein sources on BBB and neuroinflammation, which may provide an important implication for dietary guidelines for protein supplementation.

When filtrating a colloidal dispersion, colloids accumulate at the membrane surface, thus forming a polarized layer that can ultimately turns into a deposit (= a "gel") in some extreme cases. Modeling such a filtration has always been a challenge. In recent developments, Bacchin et al. propose a model based on the analogy between permeation in a deposit and diffusion in a polarized layer [1]. The permeation flux is then given by the following equation, which is the analog of the classical Dar...

In a nematic gel we observe a low-energy buckling deformation arising from soft and semisoft elastic modes. We prepare the self-assembled gel by dissolving a coil–side-group liquid-crystalline polymer–coil copolymer in a nematic liquid crystal. The gel has long network strands and a precisely tailored structure, making it ideal for studying nematic rubber elasticity. Under polarized optical microscopy we observe a striped texture that forms when gels uniformly aligned at 35 °C are cooled to r...

In nearly all cases, electrophoresis in gels is driven via the electrolysis of water at the electrodes, where the process consumes water and produces electrochemical by-products. We have previously demonstrated that π-conjugated polymers such as poly(3,4-ethylenedioxythiophene) (PEDOT) can be placed between traditional metal electrodes and an electrolyte to mitigate electrolysis in liquid (capillary electroosmosis/electrophoresis) systems. In this report, we extend our previous result to gel electrophoresis, and show that electrodes containing PEDOT can be used with a commercial polyacrylamide gel electrophoresis system with minimal impact to the resulting gel image or the ionic transport measured during a separation.

Food gels are viscoelastic substances and several gelled products are manufactured throughout the world. The gelling agents in foods are usually polysaccharides and proteins. In food gels, the polymer molecules are not cross-linked by covalent bonds with the exception of disulphide bonds in some protein gels. Instead, the molecules are held together by a combination of weak inter-molecular forces like hydrogen bonds, electrostatic forces, Van der Waals forces, and hydrophobic interactions. Polysaccharides including hydrocolloids are strongly hydrated in aqueous medium but they tend to have less ordered structures. The mechanism of gelation depends on the nature of the gelling agent(s) and on the conditions of gel formation like the temperature, the presence of ions, the pH, and the concentration of gelling agents, etc. Characterization of gels can be performed in several ways of which rheological measurements are frequently practiced. Multi-component or mixed gel system is an important area of interest in which two or more gelling components are simultaneously used to achieve certain specific structural and functional characteristics. We here discuss about the different gels and gelling agents, the characterization of gels, and the mechanism of gelation with an emphasis on mixed or multi-component gels that would have significant commercial applications.

Full Text Available In nearly all cases, electrophoresis in gels is driven via the electrolysis of water at the electrodes, where the process consumes water and produces electrochemical by-products. We have previously demonstrated that π-conjugated polymers such as poly(3,4-ethylenedioxythiophene (PEDOT can be placed between traditional metal electrodes and an electrolyte to mitigate electrolysis in liquid (capillary electroosmosis/electrophoresis systems. In this report, we extend our previous result to gel electrophoresis, and show that electrodes containing PEDOT can be used with a commercial polyacrylamide gel electrophoresis system with minimal impact to the resulting gel image or the ionic transport measured during a separation.

Background Nowadays it is now a focus topic in schistosomiasis research to find ideal vaccine candidates and new drug targets for developing anti-schistosomiasis vaccine. We cloned a new gene, casein kinase Ⅱ beta subunit, of Schistosoma japonicum (S. japonicum) and express it in Escherichia coli (E.coli).Methods The ESTs obtained in our laboratory were analyzed by homologous searching, and a new gene was recognized. The full-length cDNA of the new gene was obtained by joining the 3'RACE PCR fragment and the EST clone. To express the new gene, the cDNA was cloned into pGEX-4T-1 vector and then transformed into E.coli JM109. The recombinant protein was analyzed by SDS-PAGE and Western-blot. Results A 908 bp cDNA was isolated from S. japonicum and identified to be casein kinase Ⅱ beta subunit gene by sequence analysis. The open reading frame of the gene encodes a protein of 217 amino acids exhibiting 75.8%, 75.8%, 73.9%, 68.2%, 51.6% identity to the amino acids sequence of the corresponding genes of Homo sapiens (H. sapiens), Xenopus laevi (X. laevi), Drosophila melanogaster (D. melanogaster), Caenorhabditis elegan (C. elegan), and Schizosaccharomyces pombe (S. promber) respectively. The predicted molecular weight of the protein was 24.921 kDa. The new cDNA sequence had been submitted to GenBank, and its accession number is AY241391. This cDNA was subcloned into the pGEX-4T-1 vector and expressed in E.coli JM109.The recombinant protein could be recognized by the S. japonicum infected rabbit serum. Conclusion The full-length cDNA sequences encoding S. japonicum casein kinase Ⅱ beta subunit were firstly sequenced, cloned, and expressed in E.coli.

Iron oxide nanoparticles have been extensively used as T2 contrast agents for liver-specific magnetic resonance imaging (MRI). The applications, however, have been limited by their mediocre magnetism and r2 relaxivity. Recent studies show that Fe5C2 nanoparticles can be prepared by high temperature thermal decomposition. The resulting nanoparticles possess strong and air stable magnetism, suggesting their potential as a novel type of T2 contrast agent. To this end, we improve the synthetic and surface modification methods of Fe5C2 nanoparticles, and investigated the impact of size and coating on their performances for liver MRI. Specifically, we prepared 5, 14, and 22 nm Fe5C2 nanoparticles and engineered their surface by: 1) ligand addition with phospholipids, 2) ligand exchange with zwitterion-dopamine-sulfonate (ZDS), and 3) protein adsorption with casein. It was found that the size and surface coating have varied levels of impact on the particles' hydrodynamic size, viability, uptake by macrophages, and r2 relaxivity. Interestingly, while phospholipid- and ZDS-coated Fe5C2 nanoparticles showed comparable r2, the casein coating led to an r2 enhancement by more than 2 fold. In particular, casein coated 22 nm Fe5C2 nanoparticle show a striking r2 of 973 mM(-1)s(-1), which is one of the highest among all of the T2 contrast agents reported to date. Small animal studies confirmed the advantage of Fe5C2 nanoparticles over iron oxide nanoparticles in inducing hypointensities on T2-weighted MR images, and the particles caused little toxicity to the host. The improvements are important for transforming Fe5C2 nanoparticles into a new class of MRI contrast agents. The observations also shed light on protein-based surface modification as a means to modulate contrast ability of magnetic nanoparticles.

The objective of the work reported in this paper was to develop a theoretical model to determine the effect of type of microfiltration (MF)-process feed, number of stages, and flux on the minimization of the MF membrane area required to produce a 95% serum protein-reduced micellar casein concentrate. The MF feed, number of stages, and flux were all factors that had an effect on the MF membrane area and should be taken into consideration when designing a MF system to produce a 95% serum protein-reduced micellar casein concentrate. Feeding the MF process with a diluted ultrafiltration retentate (DUR) diluted to the protein concentration of skim milk, as opposed to skim milk, reduced the required membrane area by 36% for a 5-stage process. When DUR was the MF feed, feed protein concentration, which depended on the number of MF stages, was optimized. The DUR protein concentration that minimized the required MF membrane area was 2.47, 3.85, 4.77, and 5.41% for a 2-, 3-, 4-, or 5-stage MF process, respectively. For a 5-stage process, increasing the protein concentration of the feed from 3.2 to 5.4% decreased the required MF membrane area by 10%. It was also found that as the number of stages increased from 2 to 5, the required MF membrane area decreased by 39%, when the MF feed was DUR at the optimal feed protein concentration. Finally, increasing the flux from 50 to 60 kg/m(2) per hour decreased the required MF membrane area by 17% when the MF feed was DUR at the optimal MF feed protein concentration. Overall, using DUR as a feed for MF could reduce the amount of MF membrane area required to make a 95% serum protein-reduced micellar casein concentrate.

Full Text Available Abstract Background The secretion of heterologous animal proteins in filamentous fungi is usually limited by bottlenecks in the vesicle-mediated secretory pathway. Results Using the secretion of bovine chymosin in Aspergillus awamori as a model, we found a drastic increase (40 to 80-fold in cells grown with casein or casein phosphopeptides (CPPs. CPPs are rich in phosphoserine, but phosphoserine itself did not increase the secretion of chymosin. The stimulatory effect is reduced about 50% using partially dephosphorylated casein and is not exerted by casamino acids. The phosphopeptides effect was not exerted at transcriptional level, but instead, it was clearly observed on the secretion of chymosin by immunodetection analysis. Proteomics studies revealed very interesting metabolic changes in response to phosphopeptides supplementation. The oxidative metabolism was reduced, since enzymes involved in fermentative processes were overrepresented. An oxygen-binding hemoglobin-like protein was overrepresented in the proteome following phosphopeptides addition. Most interestingly, the intracellular pre-protein enzymes, including pre-prochymosin, were depleted (most of them are underrepresented in the intracellular proteome after the addition of CPPs, whereas the extracellular mature form of several of these secretable proteins and cell-wall biosynthetic enzymes was greatly overrepresented in the secretome of phosphopeptides-supplemented cells. Another important 'moonlighting' protein (glyceraldehyde-3-phosphate dehydrogenase, which has been described to have vesicle fusogenic and cytoskeleton formation modulating activities, was clearly overrepresented in phosphopeptides-supplemented cells. Conclusions In summary, CPPs cause the reprogramming of cellular metabolism, which leads to massive secretion of extracellular proteins.

Full Text Available Pulsed field gel electrophoresis (PFGE, the gold standard of molecular typing methods, has a major disadvantage of an unusually long electrophoretic time. From the original protocol of 6 days, it was modified to 3 days and subsequently to a single day. We describe the procedure of stacking five to six gels one on top of another in order to increase and maximize the output in a shorter time without compromising the resolution and reproducibility. All the variables that affect pulsed field gels during electrophoresis were taken into consideration. We firstly optimized the parameters to be used and secondly determined whether stacking of five to six gels had any effect on the molecular separation during electrophoresis in comparison with a single gel run. DNA preparation, restriction, electrophoresis, staining and gel documentation was carried out based on previously published methods. Gels were analysed using BioNumerics and dice coefficient and unweighted pair group methods were used to generate dendrograms based on 1.5% tolerance values. Identical band profiles and band resolution-separation were seen in the PFGE patterns with single gel and multiple stacking gels. Cluster analysis further strengthened the fact that results from stacking gels were reproducible and comparable with a single gel run. This method of stacking gels saves time and maximizes the output at the same time. The run time for a single gel was about 28 hours, but with six stacked gels the run time was 54 hours compared with 28 x 6 = 168 hours if they were run separately as single gels thus saving time of 67.86%. Beside the big factor of saving time, stacking gels save resources (electricity, reagents, water, chemicals and working time by increasing the sample throughput in a shorter time without compromising on quality of data. But optimization of working parameters is vital depending on the PFGE system used.

Milk oligosaccharides (OS)-free complex carbohydrates-confer unique health benefits to the nursing neonate. Though human digestive enzymes cannot degrade these sugars, they provide nourishment to specific commensal microbes and act as decoys to prevent the adhesion of pathogenic micro-organisms to gastrointestinal cells. At present, the limited quantities of human milk oligosaccharides (HMO) impede research on these molecules and their potential applications in functional food formulations. Considerable progress has been made in the study of OS structures; however, the synthetic pathways leading to their synthesis in the mammary gland are poorly understood. Recent studies show that complex OS with fucose and N-acetyl neuraminic acid (key structural elements of HMO bioactivity) exist in goat milk. Polymorphisms in the CSN1S1 locus, which is responsible for synthesis of αs1-casein, affect lipid and casein micelle structure in goat milk. The present study sought to determine whether CSN1S1 polymorphisms also influence goat milk oligosaccharide (GMO) production and secretion. The GMO compositions of thirty-two goat milk samples, half of which were from genotype A/A (αs1-casein producers) and half from genotype O/O (αs1-casein non-producers), were determined with nanoflow liquid chromatography high-accuracy mass spectrometry. This study represents the most exhaustive characterization of GMO to date. A systematic and comprehensive GMO library was created, consolidating information available in the literature with the new findings. Nearly 30 GMO, 11 of which were novel, were confirmed via tandem mass spectrometric analyses. Six fucosylated OS were identified; 4 of these matched HMO compositions and three were identified for the first time in goat milk. Importantly, multivariate statistical analysis demonstrated that the OS profiles of the A/A and O/O genotype milks could be discriminated by the fucosylated OS. Quantitative analysis revealed that the goat milk samples